Cow

Mastitis and types

Mastitis
Mastitis is the term which denotes inflammatory condition of udder characterized by physical, chemical and microbiological changes in milk and pathological changes in glandular tissue of the udder. In the major milk-producing countries, 15% to 20% of cows are infected with clinical mastitis each year. In Canada and the United States, it is thought that 50% of cows have one or more infected quarters.

Types of mastitis	Characteristic symptoms or definition
Grade 1 (mild)	Changes to the milk only (color– off white/yellow/red and consistency– clotted/thickened)
Grade 2 (moderate)	Changes to the milk (color/consistency) and udder (heat, swelling, pain)
Grade 3 (severe)	Changes to the milk (color/consistency), udder (heat, swelling, pain), cow (sick cow) and or gangrenous sloughing of the teat

Causal agents

Microorganisms causing mastitis
There are a lot of microorganisms on and in udder of cows. 137 identified species and subspecies of microbes are identified that can be associated with the mammary gland of the cow. Several of them are part of the normal flora and, with few exceptions, do not cause mastitis. On the contrary, they may protect udders from infection caused by pathogenic bacteria. Several other microorganisms may, however, cause infection in the mammary glands. There are contagious microorganisms and environmental microorganisms. Infected cows are the main source of contagious microorganisms, which survive and proliferate on the skin and on teat wounds. They consist of Streptococcus agalactiae, Staphylococcus aureus and Streptococcus dysgalactiae. Environmental microorganisms (Escherischia coli and other coliforms, Streptococcus uberis) do not remain on the teat. Rather, their presence indicates a high degree of contamination of soil, bedding, and water caused by manure mainly.

Types of mastitis

Species	Main Source	Living Conditions	Propagation Factors	Symptoms	Preventive Treatment
Streptococcus agalactiae	Infected cows	Infected quarter and udder only	Using same rag for cleaning udders	Mild fever for about 24 hours, Reduced milk yield, Fore strip milk having variable clots (blood stained curd) Swelling of the lower portion of udder and teat	Wash udders after milking, reduces problem by 50% Cull infected cows
Staphylococcus aureus	Infected cows	On abnormal udder and teat, milkers, vagina, tonsils	Transmitted by hands or rags, enters during milking	Odd colored milk containing flakes and clots, Abscess formation and infection up to deep interior of udder leading to inflammation of the upper portion of udder, Patchy blue/purple discoloration of and coldness of the affected udder In chronic state, udder hardens, aqueous secretion, eventual atrophy of the quarter. Intermediate form produces granular secretion. Milk hotter than normal	Wash udders after milking, reduces problem by 50% Cull infected cows
Grade 3 (severe)	Changes to the milk (color/consistency), udder (heat, swelling, pain), cow (sick cow) and or gangrenous sloughing of the teat
Streptococcus dysgalactiae	Infected cows	Infected quarter, injuries		Reduced milk yield, Fore strip milk having variable clots (blood stained curd) Swelling of the lower portion of udder and teat High fever in serious cases	Wash udders after milking, reduces problem by 50% Cull infected cows
Streptococcus uberis	Contaminated environment	On cow’s skin, mouth, ground	Neglected udder washing, insufficient drying, lack of bedding, muddy yards	Reduced milk yield, Fore strip milk having variable clots (blood stained curd) Swelling of the lower portion of udder and teat High fever in serious cases Affects mostly dry cows and heifers.	Wash teats only, dry well with disposable paper towels for each cow Supply generous bedding
Escherischia coli	Contaminated environment	Ground, bedding (sawdust and shavings), manure, water	Dirty calving stall, lack of bedding, inadequate udder washing	Abnormal milk (watery or blood tinge and drop in milk production, Excessive udder edema Thin yellow secretions, with granular texture resembling bran. Fever more than 106 degree F along with diarrhea and dehydration	Wash teats only, dry well with disposable paper towels for each cow Supply generous bedding
Corynebacterium pyrogenes	Certain insects	Humid valleys, wooded areas		Pronounced systematic reaction due to toxins caused by bacteria. Often more than one quarter affected. They become hard, produce thick smelly secretion like cheese and difficult to eliminate. Followed by abscess that bursts, releasing creamy pus, and tissue loss.
Pseudomonas	Water source, contaminated teat dips and contaminated drugs and infusion equipment, waste feed, soil and manure	Animal skin	Direct contact and using contaminated water	Marked swelling of udder, High body temperature around 107 degree F Watery milk contain flakes or clots or blood	Cull or isolate any infected cows to stop the infection from spreading, water should be tested for Pseudomonas
Mycoplasma	Infected animals, respiratory tracts, and urogenital tracts	Infected quarters	Cow to cow during milking, improper teat sanitation, contaminated intramammary treatments, milker’s hands, and airborne transmission	Mastitis found more than one quarter sometimes in all quarters Sharp drop in milk production leads to agalactia, Watery milk with few clots to a thick colostrum like material (Sandy or flaky sedimentation in watery or serious fluid)	Maintain a closed herd, buy replacements from known Mycoplasma-free herds, bulk tank samples may be cultured periodically

Factors

Factors contributing to mastitis

Mastitis is a difficult problem because it is a disease caused by many factors. Researchers estimate that 25% of the susceptibility to udder infection is attributable to environmental factors, 20% to genetic factors, and 50% to herd management.

Diagnosis

To diagnose mastitis, it is necessary to learn how to differentiate between the signs and symptoms of the various types of mastitis infection. The key points are given below:

Check the milk: Routine examination of the milk using a filter cup to extract the first three squirts before washing (before milking) is undoubtedly the best way to diagnose mastitis. The presence of lumps, flakes, blood, etc. must be watched for. Milk that is hotter than normal may be a good indication of a Staphylococcus aureus
Palpate the udder: Particularly after milking, when it is easy to detect swelling, and fibrous, hard or injured tissue.
Be attentive: To other more evident signs such as fever, redness etc.

If these symptoms are often absent, particularly in cases of subclinical, subacute or chronic mastitis, can be detected through observation. Some tests such as Somatic cell counts (SCC), bacterial identification and the California Mastitis Test (CMT), may be useful. These tests were discussed in diagnostic services part of the website.

Prevention

Preventive measures

Milking procedures

Sanitary milking habits are important to avoid the spreading of germs or their proliferation. The purpose of hygiene is to prevent the transmission of germs from one teat to another on one cow or from one cow to another.

Udder washing

Washing the udder is hygienic and has a stimulating effect on milk flow. Adequate washing is especially important to prevent environmental mastitis, caused by coliforms and other microbes from contaminated environments. Badly washed udders contribute to the transmission of microbes rather than to their destruction.

The lowest bacterial count in milk is obtained by washing the udder in the following way:

Wet and wash the teats only using individual moist paper towels. Wetting the udder and the teats results in more bacteria getting into the milk than if only the teats are wet.
Dry with individual paper towels.

Note that teat dipping before milking in addition to drying off does not give better results than drying alone, and it increases the risks of contamination of the milk by disinfectants.

Fore milking

Removing a little milk by hand before machine milking/ hand milking serves to stimulate milk letdown and to obtain a milk sample containing a high microbial count. A filter cup is used to detect abnormal looking milk (lumpy, flakes, clots, blood etc.).

Milking sequence

It is important to milk infected cows last. If possible, milking sequence should be as follows: first lactation cows, normal cows, cows with a high cell count and then infected cows.

Other measures during milking

It is important to milk completely. With modern milkers, as long as they are well adjusted, the risks of forcing the entry of microbes at the end of milking greatly diminish. The chances of bacteria entering the udder can be reduced by diminishing the amplitude of the vacuum changes and the vacuum change speed on the teats. To do so, a good vacuum reserve and appropriate piping are necessary.

Risk of infection may be diminished if milking is finished by hand, although not realistic for an entire herd. It is important to milk twice a day, even with cows that do not produce a lot. The longer the milk remains in the udder, the greater the risk of infection. The first squirts of milk must not go on the ground as this will contaminate the bedding and floor.

Postmilking teat dipping

Using a disinfectant teat dip after each milking is a means of diminishing by about 50% the risk of infection by contagious microorganisms like Streptococcus agalactiae and Staphylococcus aureus. Teat dipping prevents populations of these microbes from developing sufficiently between milking. Teat dipping also discourages flies.

It is important that the teat dip contain up to 10% of emollients to increase the suppleness of the teats: oils, glycerine, lanoline. Healthy supple skin is an extra insurance against entry of bacteria to the udder. Staphylococcus aureus does not persist on healthy skin.

For more information please “Dry cow therapy” in Udder Health: Cattle part of the website

Cleaning equipment after milking

It is vital to clean and disinfect equipment after milking. Cider or corn vinegar and peroxide are used by some producers as alternatives to phosphoric acid and chlorine.

Hygiene and safety

Abundant bedding prevents injury to the udder, limits exposure to cold, damp floors and limits contact of the udder with manure. A minimum of 3 kg of straw per day per animal must be used (about one ton per cow per year). It is better to use a little bedding often, rather than a large quantity less often. Straw is preferred. Adding lime to the bedding can help in a stable where environmental mastitis is a problem but can also irritate the udder, the teats and the lungs when airborne.

It is important to keep away the cows from injuring their udders. The floors should not be slippery when the cows are let outdoors and there should be separators between the cows. The stable should be disinfected twice a year.

Feed

Changing feed must be done slowly. Excesses must be avoided, particularly concentrates and non-protein nitrogen feed. Calcium to phosphorous ratio (1.4 to 1.8) must be maintained, even during the dry period. Selenium and vitamin E supplements may be a good choice if the ration does not provide the required amount.

Replacement of stock

Farmer should avoid to infected animals for their farm. Animals have them tested before purchasing them and examine the udders carefully. Research reported that up to 50% of purchased cows have subclinical infections. It is better to buy only heifers (heifers generally do not have mastitis) or produce your own replacement animals. Heifers should not be suckled because this breaks the teat seal and thus facilitates entry of microorganisms that can cause mastitis at calving.

Culling

Cull those animals that are severely or affected by mastitis repeatedly. Cows with injured teats that do not heal should be put at the top of the list of animals to cull. They are up to 10 times more prone to contract mastitis. Cows that maintain a high cell count during all lactations should also be culled.

Drying off period

It is well known that mastitis often affects cows that have dried off recently. These animals should not be overfed.

In conventional agriculture, dry cows are treated with antibiotics along with the pre and post milking teat dip, one of the most effective methods to reduce incidence of mastitis. For organic agriculture, the dry cow must not be overlooked. A change in feed at drying-off is important. The following are the three steps:

Post-lactation (7 to 14 days): At post-lactation stage give a reduced diet of fibrous and poor hay to provoke rapid drop in milk flow and to stimulate the rumen. Drinking water must be drastically reduced. Some organic farmers give 4 drops of sage or menthol essential oil and charcoal two times a day to cut production at this stage.

Dry (30-90 days): At dry period diet is made up mostly of roughage with a good energy-protein and mineral balance.

Pre-lactation (7 to 14 days before calving): Moderate quantities of energy-rich concentrates are added to a balanced roughage ration.

Use of the teat dip before and after the dry period (i.e. 15 days before calving and 15 days after the dry period) may be beneficial in herds where clinical mastitis is common.

For more detail about dry cow therapy please visit “Dry cow therapy” in Udder Health: Cattle part of the website.

Treatment strategies

Treatment strategies for mastitis

It will depend on whether

The mastitis is clinical or subclinical: acute or chronic
Health status of the herd
History of mastitis

Treatment decisions during lactation should be considered only for acute clinical cases of mastitis. Chronic cases will respond poorly to antibiotic therapy. Subclinical mastitis has a tendency to self-cure. Instead, subclinical mastitis and some chronic cases can be treated in the dry period only. (Nordic guidelines for mastitis therapy, NMSM Annual Conference, 2009)

Choice of Drugs

Antibiotics
NSAID (Non- steroidal anti-inflammatory drugs)
Supportive therapy (Fluid therapy)

Indication of Antibiotics

Indication of antibiotics to treat mastitis depends on some factors:

Type of pathogen involved
Type and severity of inflammatory response
Duration of infection
Stage of lactation
Age and pregnancy status of the cows

After taking decision of using antibiotics, decisions should be made in which route antibiotics will be administered.

Route of administration

Where to target antimicrobial therapy in clinical mastitis due to different pathogens (Pyörälä 2009, modified from Erskine 2003): Some common udder pathogens location in milk, udder parenchymal tissue or as systemic infection (number of + indicate where you mainly should target your antimicrobial therapy).

Pathogen	In milk	In milk	In udder tissue	As systemic infection
Streptococcus agalactiae	+++	-	-	Intramammary
Other streptococci	+++	+	-	Intramammary
Staphylococcus aureus	+	+++	-	Intramammary and intramuscularly
Coagulase negative staphylococci	+++	-		Intramammary
Trueperella pyogenes	-	++	+++	Intramammary
Coliforms*	+	-	+++	Intramammary

*Antibiotics are often not needed, NSAIDs and other supportive therapy is first choice.

Selection of antibiotics

Selection of antibiotics for the mastitis treatment is based on

Culture and susceptibility testing
About 10-40% of clinical mastitis shows no growth of bacteria, so antibiotics need not be used.
A large proportion of Gram-negative bacteria are removed by cows own immune system, so antibiotics may be avoided.

Specific Antibiotics selection

Beta-lactamase:

Beta-lactamases are enzymes produced by bacteria that give resistance to β-lactam group of antibiotics. The major antibiotics in this group are penicillin, cephalosporin, and carbapenems. The bacteria which are capable of producing this enzyme are known as beta-lactamase-producing bacteria (BLPB). BLPB can cause multiple types of infections in humans and animals. These organisms (BPLB) indirectly release free enzyme (beta-lactam) into their surrounding environment to protect themselves & other penicillin-susceptible bacteria from penicillin therapy.

Gram-positive bacteria, β-lactamas-
First choice	Treatment with Penicillin G Supportive treatment.
Second choice	Only supportive therapy No antibiotics.
Gram-positive bacteria, β-lactamas+
First choice	Only supportive therapy, No antibiotics.
Second choice	Treatment with a β-lactamase stable antibiotic (cloxacillin, dicloxacillin, flucloxacillin, methicillin) Supportive treatment.
Gram-negative bacteria (E. coli)
First choice	Only supportive therapy No antibiotics.
Second choice	Treatment with an antibiotic effective against gram-negative bacteria Supportive treatment.
Gram-negative bacteria (Klebsiella spp.)
First choice	Treatment with med Quinolones Supportive treatment should be added.
Second choice	Treatment with Trimetoprim Sulfa Supportive treatment.
Organisms	Length
Staphylococcus aureus	5 days If not cured after 5 days, the treatment could be extended for another 1 – 2 days.
Streptococcus uberis	5 days
Other gram-positive bacteria	3 – 5 days according to severity and herd.
Klebsiella spp.	3 days
Other gram-negative bacteria	3 days
No growth	Stop the antibiotic treatment.

Note: Farmers should complete the full length of the treatment. They should not stop before if the cow gets better and they should not extend too long either. It will take time for the clinical symptoms to resolve even if the bacteria are gone. If the animal does not respond to the treatment with correct antibiotic and dose, the diagnosis should be confirmed at a certified mastitis laboratory as soon as possible.

Guidelines for antibiotic use for treatment of mastitis

Milkers should be trained to detect mastitis cases early and collect milk samples aseptically. Samples should be sent to laboratory to get a basic diagnosis (no growth, Gram positive or Gram negative) to guide therapy. Cows with mild or moderate infection of clinical mastitis should be isolated and milk discarded for 24 hours until culture results are known. If the farmer wishes to initiate treatment can be given but the treatment can be modified after culture results are known
Treatments should be administered only after a person who works closely with the local veterinarian, has reviewed the medical history of the cow and evaluated the chances for therapeutic success. Cows that are third lactation or greater, have a history of previous clinical cases, or have a history of chronically high SCC are often poor candidates for routine therapy. Treatment decisions for these cows should be based on culture results. In other instances, culling, drying off the affected quarter, or extended duration therapy may be preferred
Only acute clinical mastitis cases should be chosen for treatment decision
Sub-clinical mastitis in general has high self-cure or too low cure rate in proportion to the treatment costs during lactation
Sub-clinical mastitis should be treated during the dry period
Results of treatments should be monitored. The rate of recurrence (within 60 to 90 days) and SCC reduction (by 60 days) should be recorded

Supportive treatment

Milking frequency should be increased using the milking machine or in hand milking in cases. Milk let down can be influenced by oxytocin I/M injection. Intravenous or oral fluid therapy can be administered. 1-2ml Oxytocin injection can be administered to milk let down in cows (Oxin®, Oxitocina Diana®, Uni-Oxytocin®). Oxytocin should mainly be used for mastitis caused by Gram-negatives (can even have a negative effect on mastitis caused by streptococci). The animal can be milked after 5-10 minutes. Caution is a must during prepartum usage of oxytocin. For prepartum usage cervix must be dilated naturally or artificially by estrogen or prostaglandin. Antihistamines should be used in case of gangrenous mastitis. Anti-inflammatory drugs can be used in swollen painful udder and affected general condition or as a treatment of endotoxins released by e.g. Gram-negative pathogens. NSAIDs are mainly recommended for moderate and severe cases of mastitis. Intravenous or oral fluid therapy can be administered if the cow is dehydrated.

Prognosis evaluation:

If replacement animals are in stock, it should be used for culling as an alternative for doubtful prognosis. Cost for culling and effective treatment costs should be comparatively evaluated.

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Advantages

Dry cow therapy (DCT)

Dry cow therapy (DCT) is the treatment of cows at the end of lactation with a long acting antibiotic preparation with or without a teat sealant. This is to treat for any intra-mammary infections (IMI) contracted during lactation and provides protection against new infections during the dry period.

Use of dry cow treatment is one component of an effective mastitis control program that should also include: proper milking procedures using properly functioning milking equipment; dipping teats immediately after milking with a product known to be safe and effective; good udder hygiene between milking; keeping accurate records of clinical mastitis and somatic cell counts on individual cows to assist in making management decisions; treating all clinical cases of mastitis promptly and appropriately; and culling cows with chronic mastitis.

Advantages of DCT:

🡪Curing Existing Infections

The most effective time to treat subclinical udder infections is at drying off. Dry cow therapy has the following advantages over lactation therapy:

The cure rate is higher than that achieved by treatment during lactation, particularly for Staphylococcus aureus.
A much higher dose of antibiotic can be used safely.
Retention time of the antibiotic in the udder is longer.
Tissue damaged by mastitis may be regenerated before freshening.
The risk of contaminating milk with drug residues is reduced when the milk withholding time after calving is properly observed.

🡪Preventing New Infections

The risk of new intramammary infection is greatest during the early and later portions of the dry period. Most dry cow treatments provide sufficient protection after drying off so that:

The frequency of new infections during the dry period is reduced.
The incidence of clinical mastitis at freshening may be reduced.

Types of dry cow therapy:

Blanket dry cow therapy:

Blanket dry cow therapy is the process of treating every quarter of every cow with antibiotics at drying off, independent of their infectious stage.

Use of antibiotics in animals differs between countries. In most countries blanket DCT is recommended. Other countries, specifically the Nordic countries have been more restrictive. In those countries the restricted use of dry cow antibiotics generally did not lead to unacceptable high incidence rates of clinical mastitis.

Estimated use of blanket DCT:

USA 72%
Netherlands 76%
Canada 88%

In the Nordic countries the quantity of antibiotics used is much lower:

Norway 1-2%
Denmark 6-7%
Finland 20%
Sweden 25%

This blanket approach will reach all infected quarters, is more effective than selective treatment in preventing new infections early in the dry period, and does not require laboratory screening procedures to decide which cows and quarters to treat. However, selective treatment may fail to reach 20 to 40 percent of infected quarters in a herd. Also, uninfected quarters not treated at drying off are more likely than treated quarters to become infected during the dry period.

Selective dry Cow Therapy (SDCT)

Selective dry Cow Therapy (SDCT) is a process of selecting cows to treat with antibiotics at drying off on an individual cow needs basis. This can be done by carefully planning and using records to select the appropriate cows that need treatment and simply using teat sealants on their own with the rest of the herd. Selective DCT can be very successful, but only if the cows are chosen correctly and sealer applied hygienically.

Selective DCT is only suitable for herds where:

Bulk tank SCC is less than 200,000-250,000 cells/ml all year
Individual somatic cell count data available
At least four milk recording results are available
Milk culturing tests have been carried out
Clinical cases of mastitis have been recorded
Mastitis is under control (Low incidence, <2%) in the herd
Teat sealants used in all cows
Dry cow plan, especially around environmental hygiene in the dry period should be performed

Otherwise, use blanket DCT

Selective DCT is only suitable for cows where:

Individual SCC <200,000 for the last 3 milk consecutive tests
No clinical cases of mastitis detected in previous lactation

Hygiene at drying-off is even more important if the cow is being given sealer only, with no antibiotic tube. Treatment with teat sealant to prevent new infections during the dry period. It is essential this is administered using a very hygienic procedure to ensure infection is not introduced with the teat sealant.

The advantages of SDCT

Save money as not all cows get DCT
Only cows that need antibiotics are treated with them
Reduced antibiotic usage on farm
Reduced risk of antimicrobial resistance on farm
Reduce the risk of severe toxic mastitis in early lactation. There is evidence that unnecessary antibiotic DCT in cows that are uninfected at dry-off can increase the risk of severe mastitis in early lactation by removing the commensal bacteria from the udder.

Early dry period

Drying off and the early dry period (7-8 weeks prior to calving)

Reducing the grain ration and sudden cessation of milking is the recommended practice for drying off cows. High producing cows should be taken off concentrates two weeks prior to dry off to help reduce production. Intermittent milking along with a decrease in the energy concentration of the ration can be used as a method to achieve the target yield. Cows should be observed closely for two weeks after drying off to ensure udders are involuting (not swollen or inflamed) properly. Udders with swollen quarters should be examined for mastitis. Cows showing visible signs of illness should be provided supportive therapy; however, re-infusion of antibiotics into the mammary gland is not recommended. Supportive therapy may include intramuscular or intravenous administration of antibiotic and/ or anti-inflammatory compounds. In severe cases electrolyte therapy may be warranted. A veterinarian should be consulted for advice on a proper treatment procedure.

Late dry period

Late dry period (2-3 weeks prior to calving)

While dry cow treatment is beneficial in preventing new infections during the early dry period, the udder is vulnerable to new infections during the last two to three weeks of the dry period when dry cow therapy is no longer effective. Special attention must be given to springing cows and heifers. These animals must be kept clean and dry if mastitis is to be avoided during early lactation. Weather permitting, a clean grassy lot is an ideal calving area. Stalls with clean dry bedding, preferably straw or inorganic bedding, are recommended during inclement weather.

Recent studies on the use of persistent barrier teat dips starting 10 to 14 days pre-partum may prove a viable management option for reducing new intramammary infections at calving. Studies indicate barrier dips persisting greater than three days may result in up to a 50 percent reduction in total, major pathogen and environmental streptococcal infections at calving in cows and heifers. This practice may be particularly beneficial in herds experiencing high rates of mastitis in early lactation or when environmental conditions are less than ideal.

Factors

Factors to consider from dry off to calving:

Calving date - cows should get a minimum dry period of two months before the next lactation
Withhold period of anthelmintic or dry cow antibiotic treatment
Low volume - dry off cows as soon as their daily milk production drops to 9L
Dry off cows abruptly, as soon as production reaches 9 liters per day
Dry off cows in small batches (e.g. one row of the parlor at a time)
Don’t dry off cows on an empty stomach, have some breakfast first
Don’t leave cows in roadways or yards after drying off, leave in an area that is clean for lying on.

An extended dry period should be considered for:

High SCC cows – dry off early to improve mastitis cure rates (>9 weeks)
Low Body Condition Score cows -especially first calves. Cows should be dried off at BCS 3.0-3.25, which is the condition in which they should calve down

Example	Length of dry period
	BCS 2.5	BCS 2.75	BCS >3.0
	(12-14 weeks dry)	(8-10 weeks dry)	(8 weeks dry)
Expected calving date	Dry off date	Dry off date	Dry off date
Feb 1	Nov 30	Nov 16	Nov 2
Feb 8	Dec 7	Nov 23	Nov 9
Feb 15	Dec 14	Nov 30	Nov 16
Feb 22	Dec 21	Dec 7	Nov 23
Mar 1	Dec 28	Dec 14	Nov 30

Total procedure

Total procedure from drying-off to calving

During milking, mark and draft out the group of cows to be dried off
Finish milking, clean out the shed
Use a new pair of gloves and keep them clean while drying off cows
Mark the cows again with a different color to indicate that they are now being treated; it is important to do this before they are treated
Disinfect the teats with teat dip or spray and then wipe off
Then sterilize the teat ends with cotton wool or medicated wipes
Administer the antibiotic tube and massage into the udder
When inserting the sealer, pinch the base of the teat so that the sealer remains in the teat canal and do not massage up into the udder
Record the treatment details: animal numbers, date, product used and withholding time

Anti-microbials procedure

Dry cow antimicrobials:

Dry cow products contain high levels of one or more antibiotics in a slow-release base which will maintain therapeutic levels in the dry udder for a significant length of time. Further, they have been tested in field studies, meet the guidelines of the regulators, and are guaranteed to be prepared aseptically.

All syringes used must be for single infusion only. Unapproved products and nonstandard methods may lead to the infusion product becoming contaminated during mixing and through multiple use, and they may spread resistant organisms. Products used for dry cow therapy should be stored in accordance with good dairy farming practices and discarded when the expiration date is reached.

Most dry cow therapy products are designed to eliminate existing infections by Gram positive bacteria, particularly Staphylococcus aureus and streptococcal infections at drying off, and to prevent new S. aureus and streptococcal infections in the early dry period. Many producers have already eliminated Streptococcus agalactiae and dramatically reduced the level of S. aureus infection in their herds.

Continued use of dry cow treatment will help to maintain a good herd udder health status. In many herds and especially where dairy cattle confinement has become more intense, a higher percentage of new infections during the dry period are caused by environmental bacteria. Most dry cow therapy products are reasonably effective against environmental streptococci, especially Streptococcus uberis, but they lack activity against Gram-negative environmental bacteria, especially the coliforms. The length of effective protection varies between products, often according to the type of antibiotic or the dose. In Europe and Australia, dry cow products providing protection up to 54 days are available. The herd veterinarian should be consulted to determine which dry cow product should be used.

Dry cow products

Product name	Composition	Action	Dose	Withdrawal period	Company
Bovaclox™ Dry Cow	Cloxacillin 500mg, Ampicillin 250mg v	Staphylococcus aureus, Streptococcus uberis, S. agalactiae, S. dysgalactactiae, Corynebacterium spp. and E. coli.	1 syringe per quarter immediately after final milking	Milk: from the first 8 milkings after calving must be discarded Meat: 30 Days	MSD Animal Health
Cepravin® Dry Cow/ Cefshort®	Cephalonium 250mg	Staphylococcus aureus, coagulase negative Staphylococci (CNS), Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae,Corynebacterium and Escherichia coli	1 syringe per quarter immediately after final milking	Milk: from the first 8 milkings after calving must be discarded Meat: 30 days	MSD Animal Health/ Zoetis
Cefa-Safe®	Cephapirin benzathine 300mg	Bacteria sensitive to cephapirin	1 syringe per quarter immediately after final milking	Milk: from the first 8 milkings after calving must be discarded Meat: 30 days	MSD Animal Health
Quartermaster®	Procaine Penicillin G 1 million IU and Dihydrostreptomycin Sulfate 1gm	Staphylococcus aureus	1 syringe per quarter immediately after final milking.	Milk: 96 hours Meat: 60 days	Zoetis
SPECTRAMAST®DC	Ceftiofur hydrochloride 500 mg	Staphylococcus aureus, Streptococcus dysgalactiae and Streptococcus uberis.	Infuse 1 syringe into each affected quarter at the time of dry off	Milk: 0 hour Meat: 16 days	Zoetis
Cefa-Dri® ToMORROW®	cephapirin benzathine-300 mg	streptococcus agalactiae and Staphylococcus aureus	Infuse a 10 mL syringe once each quarter has been completely milked out.	Milk: 72 hour Meat: 42 days	Zoetis, Boehringer Ingelheim Vetmedica, Inc
Cefimam DC ®	Cefquinome Sulfate	Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus agalactiae, Staphylococcus aureus, coagulase negative staphylococci.	Infuse 1 syringe into each affected quarter at the time of dry off.	Meat and offal: 2 days Milk: 1 day	Norbrook
Ubro Red®	aminoglycoside, a penicillin ester and a penicillin	Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Trueperella pyogenes, Escherichia coli and Klebsiella spp.	Infuse 1 syringe into each affected quarter at the time of dry off.	Milk: 84 hours Meat: 28 days	Boehringer Ingelheim Vetmedica, Inc
ALBADRY PLUS®	Penicillin G procaine 200,000 IU and Novobiocon 400mg	Staphylococcus aureus, Streptococcus agalactiae	Infuse 1 syringe into each affected quarter at the time of dry off	Milk: 72 hours Meat: 30 days	Zoetis
Orbenin® -DC Dry-Clox®	Cloxacillin 500 mg	Staphylococcus aureus, Streptococcus agalactiae	1 tube in each teat	Milk: 0 hours Meat: 28 days	Zoetis

Infusion procedures

The teats must be cleaned and sanitized carefully before infusion. Without proper preparation, organisms present on the teat end may be forced into the udder and result in a severe infection, especially if Gram-negative bacteria are introduced.

The best procedure is to follow these easy steps:

Clean and dry teats.
Dip teats in an effective germicidal product. Allow 30 seconds contact time before wiping teats with an individual disposable towel.
Thoroughly clean and disinfect each teat end, paying particular care to the teat orifice, by scrubbing with a cotton swab soaked in 70 percent alcohol. Use a separate piece of cotton for each teat.
Prepare teats on the far side of the udder first, followed by teats on the near side. (Teats may be cleaned and infused individually, if necessary.)
Treat quarters in reverse order; near side first, far side last.
Insert only the tip of the cannula into the teat end and express all of the contents. Do not allow the sterile cannula to touch anything prior to infusion.
Do not massage the teats to disperse the product.
Dip teats in an effective germicidal product after treatment.
Identify treated cows and remove them from the milking herd to prevent antibiotics from entering the milk supply.

Withdrawal periods

Due to the long acting nature of the antibiotics used, there is a prolonged milk withdrawal period associated with their use. It is important that if the cow calves early that the instructions are appropriately followed for each antibiotic product. Failure to do so may lead to an antibiotic residue failure. Milk can be tested using an inhibitory substance test.

Recommended Dietary nutrient concentrations (DM basis) suggested for pregnant non-lactating cow

Nutrients	Unit	Early dry	Close up dry
Net energy	MCal/kg	1.10-1.20	1.45-1.55
Crude protein (CP)	% DM	12-13	13-14
Soluble CP	% CP	40-50	35-45
Degradable CP	% CP	65-70	62-67
Undegradable CP	% CP	30-35	33-38
Acid Detergent Fiber	% DM	35-40	30-35
Neutral Detergent
Neutral Detergent
Fiber	% DM	50-65	40-55
Calcium	% DM	0.31-0.35	0.36-0.41
Phosphorus	% DM	0.19-0.21	0.22-0.25
Magnesium	% DM	0.18-0.20	0.22-0.25
Sodium	% DM	0.10-0.13	0.12-0.15
Chloride	% DM	0.20-0.22	0.24-0.26
Sulfur	% DM	0.16-0.18	0.19-0.21
Cobalt	PPM	0.10	0.12
Copper	PPM	10-15	12-18
Iron	PPM	50	60
Iodine	PPM	0.60	0.70
Manganese	PPM	40	50
Zinc	PPM	40	50
Selenium	PPM	0.30	0.30
Vitamin A	IU/kg	4000	4700
Vitamin D	IU/kg	1200	1400
Vitamin E	IU/kg	25	30

*Early dry period is defined as 4 - 6 weeks after dry off with suggested nutrient densities based on a dry matter intake of 1.9-2.1 % of body weight.

*Close-up dry period is defined as 2 - 4 weeks prior to calving with suggested nutrient densities based on a dry matter intake of 1.6-1.8% of body

Chronic high SCC cows

In cows with a history of a high SCC during the previous lactation, there is an opportunity at drying off to use a combination of antibiotic treatments (both intramammary preparation with parenteral antibiotics) to try to maximise the probability of bacterial cure. The choice of antibiotics used should be based on milk cultures or, alternatively, farm pathogens known to be present.

Heifers

Sometimes farms have problems with mastitis (clinical or subclinical) after heifers have calved. This is due to their environmental management pre calving. Some farms have had success in using internal teat sealants on heifers to prevent this occurring. Alternatively a barrier teat dip may be applied on a regular (perhaps weekly) basis prior to calving.

Teat sealant

Internal teat sealant infusions

The National Mastititis Council’s Recommended Mastitis Control Program suggests using a teat sealant on dry cows exposed to a high level of environmental pathogens. An effective dry cow teat sealant will provides an external physical barrier for the teat orifice during critical times in the dry period.

Sealing of the teat canal by the natural keratin plug that forms during the dry period is the primary natural component protecting against new intramammary infection in the late dry period. Potential damage to that protection is one reason why repeated infusions are not recommended. It has been documented that a significant proportion of quarters experience long delays or outright failure to form a complete keratin plug during the dry period, placing these quarters at increased risk for experiencing new mastitis infections. One study has reported that this risk is increased in cows producing high levels of milk at dry off.

One method of supplementing the teat’s defenses throughout the entire dry period is by use of an internal teat sealant. An artificial internal sealant is available for use alone or in combination with an antibiotic infusion. This product has no antimicrobial activity and, therefore, is recommended for use alone only in the uninfected udder. Otherwise, internal teat sealants should be used in conjunction with dry cow antibiotic therapy. When used alone in uninfected quarters, this product has been shown to prevent significantly more new infections than using no treatment at all, and it has been shown to have equal, if not better, efficacy in preventing new infections as compared to using antibiotic alone.

In the infected udder, or when the infection status is unknown, an antibiotic infusion is recommended. This may be accompanied by teat sealant and may be especially valuable for the longer dry period. Using the internal sealant in combination with an antibiotic prevents significantly more new dry period infections than using antibiotic alone.

While internal teat sealants are most commonly used in combination with intramammary antibiotics in North America, they are also approved for combination use in most European countries. The teat sealant meets all requirements for protection of the nonlactating gland for organic herds, but this is also dependent on individual countries’ requirements. It is paramount that the very best hygienic practices are adopted when infusing the teat sealant to prevent contamination of the mammary gland.

Internal teat sealants

Products name	Compositions	Dose	Companies
Orbeseal®	Bismuth subnitrate	One syringe for each udder quarter	Zoetis
Sureseal® Teat Sealer (Singles)	Bismuth subnitrate	One syringe for each udder quarter	Norbrook
Teatseal®	65% bismuth sub nitrate in a paraffin base	One syringe for each udder quarter	Zoetis

After using dry cow treatment, the following steps to infuse internal sealant:
1. Teats should be clean and dry. If teats are not clean, CAREFULLY wash and dry them before disinfection.
2. Using an alcohol pad, physically clean the end of the teat to remove any contaminated skin, dirt or manure. Repeat until the pad remains clean.
3. Disinfect the far teats before the near teats to avoid accidental contamination of previously disinfected teats.
4. Insert the internal sealant syringe nozzle into the teat canal, grasp the base of the teat near the udder attachment with two fingers press firmly together and slowly inject all contents. Use one complete syringe per quarter. Do not massage. Internal sealant must remain in the teat to be effective.
5. Insert internal sealant into the nearest teats first, to minimize contamination of teats that have not been treated.
6. After inserting internal sealant, mark the cow so others can tell she has been dried off. Then dip each teat with a quality teat dip.
Internal sealant requires no milk or pre-slaughter withdrawal. Follow the withdrawal rules described on the label, when internal sealant is used in conjunction with a dry-cow treatment program.

Remove internal sealant, program flowing steps:
1. To effectively strip internal sealant, be sure to grab the top of the teat-where it meets the udder-and work all the way down. Don’t grab the middle of the teat, squeeze and work down. This will only clear the bottom half of the teat. Strip the entire quarter by starting at the top and working all the way down.
2. Strip aggressively-10 to 12 times per quarter-for the first 4 days post-freshening. This helps to ensure that you're removing the plug and all internal sealant particles. Do not remove internal sealant by action of the milking machine.
3. Milk into a bucket for the first 3 to 4 days post-freshening. This will help to remove any remaining internal sealant particles.
2 External teat sealants

External teat sealants are in the form of a barrier film that is applied to the teat skin as a dip. Its duration of action is thought to be approximately 7 days. However, it is considered to be a less effective preventative of new infections during the dry period than the internal teat sealants and therefore is less widely used. The efficacy can be increased by regular application during the dry period although this is likely to be too inconvenient for the farmer to consider.

Teat dipping

Dipping teats with a disinfectant is considered one of the most important steps in the prevention of new mastitis infections. When the practice of teat dipping is employed, the rate of new infections during lactation can be reduced approximately 50 percent within one year. After a two-year period, up to 75 percent of the infections can be prevented. If teat dipping is discontinued, the infection rate increases rapidly.

Dipping teats both pre- and post-milking with a disinfectant containing chlorous acid and chlorine dioxide reduced Staph aureus infections by nearly 70% and Strep uberis infections by 65% when compared with post-dipping alone.

Pre-dipping:

Pre-dipping means applying a quick-acting disinfectant just before milking to reduce the population of mastitis-causing bacteria on teat skin especially in the region of the external teat orifice. The major effect of pre-milking teat disinfection is therefore against those environmental micro-organisms which cause mastitis. Pre-milking disinfection is not aimed at improving teat condition.

Pre-dipping reduces new environmental streptococcal infections and Escherichia coli by as much as 50%. Pre-dipping teats should be considered if there are high numbers of mastitis cases due to environmental bacteria (> 5 per 100 cows per month), or cows at high risk periods such as in the first week after calving.

Pre- dipping can be summarised as: “Strip, Dip, Dry and Apply”. That is:

Strip: Wipe loose dirt from each teat and pre-strip foremilk;
Dip: Pre-dip and wait for at least 15 sec, preferably 20-30 sec;
Dry: Wipe each teat with a single-use paper towel;
Apply: Apply teatcups 60-90 sec after the first touch of each udder.

Post- dipping:

Dipping the teats immediately after milking has been practised for many years. The aim of post-milking dipping is to remove any contagious mastitis-causing pathogens that may have been deposited on the teat surface and maintain the healthy teat skin.

omplex Chemical & Pharmaceutical (ethyleneoxy) ethanol-iodine complex

Product	Manufacturer	Active Ingredients
			Indications
			Pre-dip	Post-dip
3M Lauricare Moisturizing Teat Dip Concentrate	3M Canada Company	Caprylic/capric acid monoesters, glycerol monolaurate		✔
4XLA	Ecolab Co.	Glycerin, lactic acid, sodium chlorite	✔	✔
Agro Blue	Westagro Canada	Glycerin, chlorhexidine acetate		✔
Auto Chlorhex Canada	Agrisan Specialty Chemical & Pharmaceutical	Glycerin, chlorhexidine acetate		✔
Blue Velvet Teat Dip	London Dairy Supply Ltd	Glycerin, chlorhexidine acetate		✔
H-30 Teat Dip 0.55%	Ostrem Chemical Co Ltd.	Glycerin, chlorhexidine acetate		✔
HX Dip (Post Milking Teat Dip)	Ecolab Co.	Glycerin, chlorhexidine acetate		✔
Della-Blue	DeLaval Inc	Glycerin, chlorhexidine acetate		✔
AgroB B-550	GEA Farm Technologies Inc.	Chlorhexidine acetate		✔
Apprehend	GEA Farm Technologies Inc.	Chlorhexidine acetate		✔
Teat Dip w Chlorhexidine Liq 0.5%	Western Chemical Inc.	Chlorhexidine acetate		✔
Theratec Plus (Pre and Post Antiseptic Teat Dip with Added Emollient)	GEA Farm Technologies Inc.	Iodine (nonylphenoxypoly (ethyleneoxy) ethanol-iodine complex)	✔	✔
Theratec Pre and Post Sanitizing Teat Dip	GEA Farm Technologies Inc.	Iodine (nonylphenoxypoly (ethyleneoxy) ethanol-iodine complex)
Bovitec	Agrisan Specialty Chemical & Pharmaceutical	Iodine (nonylphenoxypoly (ethyleneoxy) ethanol-iodine complex)
Della-One	Delaval Inc.	Glycerin, iodine (polyethoxy polypropoxy polyethoxy ethanol - iodine complex)		✔
Della-Pretech Plus	DeLaval Inc.	Glycerin, iodine (polyethoxy polypropoxy polyethoxy ethanol - iodine complex)	✔	✔
Della-Pro	DeLaval Inc.	Iodine (hydriodic acid – iodine complex)	✔	✔
Della-Soft Act	Delaval Inc.	Glycerin, iodine		✔
Iodip 5	Agrisan Specialty Chemical & Pharmaceutical	Nonylphenoxypoly (ethyleneoxy) ethanol-iodinec	✔	✔
Armor	GEA Farm Technologies Inc.	Iodine		✔
Oxy-Gard Sanitizing Teat Dip	Ecolab Co.	Hydrogen peroxide, lactic acid, sodium linear alkylate sulfonate	✔	✔
Preodyne	Agrisan Specialty	Nonylphenoxypoly	✔	✔
B-30	GEA Farm Technologies Inc.	Alkyl phenol polyglycol ether iodine complex, glycerin, nonyl phenoxypoly ethoxy ethanol-iodine, polyoxyethylene glycol iodine complex		✔
Barricade	Ecolab Co.	Lanolin, nonyl phenoxypoly ethoxy ethanol-iodine complex		✔

ReferenceBiggs A. 2009. Mastitis in Cattle, 1st Edition. The Crowood Press Ltd.Blowey R, Edmondson P. 2010. Mastitis Control in Dairy Herds, 2nd Edition. CABI International.Bradley AJ, Green MJ. 2004. The importance of the nonlactating period in the epidemiology of intramammary infection and strategies for prevention. Veterinary Clinics of North America: Food Animal Practice. 20:547-568
Dingwell RT, Timms LL, Sargeant JM, Kelton DF, Schukken YH and Leslie KE. 2003. The association of teat canal closure and other risk factors for new dry period intramammary infections. National Mastitis Council 42nd Annual Meeting Proceedings, 298-299.

http://www.msd-animal-health.com/

http://www.nmconline.org (2006)

http://www.osuextra.com

https://en.wikivet.net/Dry_Cow_Therapy

https://www.bi-vetmedica.com/

https://www.farmacy.co.uk/categories/91-dry-cow-tube

https://www.zoetis.com/

Orbeseal® is a registered trademark and Prevent New Infections, Naturally™ is a trademark of Zoetis Inc. Zoetis Inc ORB03019

www.mbmvetgroup.co.uk

www.bimeda.co.uk

http://www.milkproduction.com/Library/Editorial-articles/Dry-cow-therapy--does-it-still-deserve-a-blanket-recommendation/

Halasa, T., et al. "Meta-analysis of dry cow management for dairy cattle. Part 1. Protection against new intramammary infections." Journal of Dairy Science 92.7 (2009): 3134-3149.

http://www.nadis.org.uk/disease-a-z/cattle/mastitis/mastitis-part-7-teat-disinfection/

Oliver SP, Lewis MJ, Ingle TL, Gillespie BE, Matthews KR. 1993. Prevention of bovine mastitis by a premilking disinfectant containing chlorous acid and chlorine dioxide. Journal of Dairy Science. 76:287-292

Pankey JW, Wildman EE, Dreschler PA, Hogan JS. 1987. Field trial evaluation of premilking disinfection. Journal of Dairy Science. 70:867-872

Smith KL, Hogan JS. 1997. Risk factors for environmental streptococcal intramammary infections. In: Proceedings of the symposium on udder health management for environmental streptococci, Ontario Veterinary College, Canada. 42-50

Blowey R, Edmondson P. 1995. Teat disinfection. In: Mastitis control in dairy herds, Chapter 7, Farming Press Books, Ipswich, United Kingdom. 93-102

Biggs A. 2009. Mastitis in Cattle, 1st Edition. The Crowood Press Ltd

Blowey R, Edmondson P. 2010. Mastitis Control in Dairy Herds, 2nd Edition. CABI International

Van Saun RJ. 1991. Dry cow nutrition: the key to improving fresh cow performance. Veterinary clinics of North America: Food animal practice, 7(2), 599-620

National Research Council. 2001. Nutrient requirements of dairy cattle: 2001. National Academies Press

Buffalo

History

The domesticated water buffalo is a potential livestock species that supplies humankind with milk, meat, and draught power. This species is reared in almost 77 countries covering Asia, Europe, Africa, and America. Two types of domestic Asian water buffalo descended from different wild Asian water buffalo (Bubalus arnee) populations some 900 thousand years ago which later spread into other geographical regions. It is believed that the river buffalo (B. bubalis bubalis) was domesticated in the western region of the Indian subcontinent (6300 years ago) and spread to Egypt, the Balkans, Greece, and Italy. On the other hand, swamp-type (B. bubalis carabanesis) buffalo were domesticated near the border of the China and Indochina region 3000-7000 years ago and dispersed through Southeast Asia, Assam and Bangladesh, and China

Distribution

Because of the high-quality products and their adaptability to harsh environmental conditions, interest in buffalo production has increased. Buffaloes are found in India, Pakistan, Bangladesh, Iraq, Iran, Nepal, Myanmar, the Philippines, Turkey, Italy, China, Thailand, Brazil, and Egypt. The number of buffaloes worldwide was reported to be 206 million in 2018. Asia has the majority of the buffaloes (97.3%), followed by Africa (1.7%), America (0.7%), Europe (0.2%), and Oceania (0%). Table-1 shows that India has the largest buffalo population of all countries. This buffalo population contributes 55% of the total milk in the country. Some of the best and most productive buffalo breeds originated from India, such as Murrah, Nili-Ravi, Surti, and Jafarabadi.

Buffalo population in different countries

Countries	Buffalo population (in million)
India	114.15
Pakistan	38.84
China	27.11
Brazil	1.39
Italy	0.40
Malaysia	0.11
Sri Lanka	0.30
Thailand	1.25
Vietnam	2.42
Philippines	2.88
Myanmar	3.79
Nepal	5.27
Bangladesh	1.48

Figure 1.2: World buffalo distribution map (Credit: Minervino et al., 2020)

Types of buffalo breeds

The two major types of wild buffaloes are Asian water buffalo (Bubalus arnee) and African Buffalo (Syncerus caffer). The two subspecies of Asian water buffalo are: the river buffalo (B. bubalis bubalis), most found in the Indian subcontinent and some European and American countries, and the swamp type buffalo (B. bubalis carabanesis,) mainly found in Bangladesh, China, Southeast Asia, Australia, and north-eastern states of India.

Points	River type buffalo	Swamp type buffalo
Distribution	India, Pakistan, Bangladesh, some European and American countries	Bangladesh, China, Thailand, the Philippines, Indonesia, Australia, the North-eastern states of India, etc.
Chromosome no.	50 chromosomes	48 chromosomes
Horn	Relatively straight pale-colored horns or curled.	Generally curved, massive backswept horns.
Color	Black or dark grey without white markings on the leg and face, mainly	Slate gray with white chevron (one or two white stripes on the throat), socks, and tail tip.
Adult weight	450 - 1000 kg	325 - 450 kg
Production	Milk production is higher than swamp-type buffalo	The females yield up to 600 kg of milk per lactation
Utility	Mainly for milk production and suitable for meat and draught purposes as well	They are mainly used for draught purposes and are poor milk producers.
Figure

Some important buffalo breeds of Asia and Europe are-

Traits	Picture
Murrah ● Origin: The center of Haryana and spread across the country. ● Distribution: Different states of India, Brazil, Bulgaria, and many Asian countries. ● Skin and hair color: Jet black with a white switch in the tail. ● Horn: Short and tightly curled. ● Body weight: o Bull: 750 kg o Cow: 650 kg ● Milk production: 1,800 kg ● Lactation period: 305 days ● Milk fat: 7.2%	Fig: Murrah Bull (Credit: Indiamart)
Nili-Ravi ● Origin: Around the Ravi River, India. ● Distribution: Ferozpur district, Punjab, and Sahiwal (Pakistan). ● Body color: Black with white spots in extremities and walled eyes. ● Horns are less curled compared to Murrah buffalo. ● Udder: Well-shaped and extends well forward to the naval flaps. ● Body weight: o Bull: 700 kg o Cow: 600 kg ● Milk production: 2,000 kg ● Lactation period: 305 days ● Milk fat: 6.5%	Fig: Nili-Ravi bull(Credit: Hisarbovine)
Kundi ● Origin: Indus Valley ● Distribution: Sindh region of south Pakistan ● Body color: Jet black ● The horn is short with a broad base and tapers upward and inward ● Udder is well developed with prominent milk veins, and teats are squarely placed ● Body weight: o Bull: 700 kg o Cow: 600 kg ● Milk production: 2,000 kg ● Lactation period: 320 days ● Milk fat: 7%	Fig: Kundi
Surti ● Origin: Southwestern part of Gujarat ● Distribution: Gujarat and Rajasthan district of India. ● Black coat color with black or reddish skin and two white chevrons on the chest ● Characterized by white markings on the forehead, legs, and tail tips. ● Horns are flat, medium length, and directed downward and backward ● Body weight: o Bull:700 kg o Cow: 550-650 kg ● Milk production: 2,090 kg ● Lactation period: 350 days ● Milk fat: 6.6-8.1%	Fig: Surti (Credit: TradeIndia)
Jafarabadi ● Origin: Gujarat, India. ● The breeding tract is Gir forests, Kutch, and Jamnagar districts of Gujarat in India. ● Amber-black in color with white or gray marking forehead, tail, and feet ● Horn is broad and large, directed downward and then upward. Sometimes covers the eyes. ● Body weight: o Bull: 600-1,500 kg o Cow: 700-800 kg ● Milk production: 1,800-2,700 kg ● Lactation period: 350 days ● Milk fat: 8.5%	Fig: Jafarabadi (Credit: ExportersIndia)
Mediterranean Buffalo ● Origin: IndiaInvalid source specified. ● Mainly distributed throughout Italy ● Medium body with grey hair ● Mostly black, black and brown, and dark gray ● The horn is flat at the bottom and faces backward, and their points face upward and inward ● Body weight: o Bull: 500-600 kg o Cow: 300-450 kg ● Milk production: 900-4,000 kg ● Lactation period: 270 days ● Milk fat: 8%	Fig: Mediterranean Buffalo
Indigenous Bangladeshi ● Found mainly in the Southwestern part of Bangladesh. ● Medium size body and black with a white spot on the forehead and tail-switch. ● The horn is medium to large and directed backward to outward. ● Body weight: 427 kg ● Milk production: 620 kg ● Milk fat: 6.8-13.2%	Fig: Indigenous Bangladeshi(Photo credit: Sanjib Chandra Nath)

Behaviour

Feeding behaviour: Buffaloes are strict grazers and usually graze during the daytime, while ruminating and sleeping are predominant activities at night. They are good converters of poor-quality roughage to high energy. These animals spend more time in rumination, less time in ingestion, and more at rest than cattle.

Heat tolerance and wallowing behaviour: Buffaloes are less tolerant to extreme heat and cold than cattle. They have thick skin with many melanin pigments, giving a black coat color. This thick and black coat color is a good heat absorber and limits heat loss from the body. Moreover, a lower density of sweat glands and hair in the skin limits heat loss. As a result, buffalo show wallowing behaviour, which is a learned behaviour. Buffaloes wallow in water during hot and cold weather. It helps to regulate their body temperature and reduce heat stress. In another way, wallowing is helpful to protect them from insects.

Social behaviour: Like many other livestock species, buffaloes live in groups of bulls, dry or pregnant cows, young stock, and calves. They communicate through different visual signals, sound, touch, and smell. The buffaloes recognize each other by smelling each other. Buffaloes will rub themselves against trees to leave a scent and defecate to mark their territory. After parturition, the dam stands up and starts to lick and sniff her calf to stimulate respiration, blood circulation, urination, and defecation. Dams quickly learn to recognize their calf and usually reject neonates from other mothers if they try to approach their udder. Both the dam and calf use vocal communication to recognize each other.

Utility of buffalo

Milk production: Buffalo is the world's second-highest milk-producing animal. But in some South Asian countries like India, Pakistan, and Nepal, the contribution of buffalo milk to total milk production is higher than cow milk. Buffalo milk has higher nutritional value, for example, more protein, vitamin, and mineral content than cow's milk. Moreover, it contains beneficial compounds that may provide antioxidant protection and improve bone and heart health. The high milk solids and fat in buffalo milk aid in preparing dairy products such as yogurt and cheese.

Meat production: Buffalo meat is very popular in most countries. About 90.5% of world buffalo meat was produced in Asia, with the more incredible amount contributed by India, with 1.6 million tons produced, followed by Pakistan with 0.93 million tons. Buffalo meat is known in various names in different countries. In some places, it is known as red beef, or buff in India and Nepal; in some countries, it is known as carabeef. It is lean and rich in protein but lower in fat, cholesterol, and calories than cattle meat. This unique nutritional profile has created a high demand among health-conscious consumers.

Draught purpose: For centuries, buffaloes have been used as draught animals as they have good muscular development. They are widely used to plough, puddle rice fields, level land, cultivate field crops, haul carts, sleds, shallow-draft boats, etc. The strong large feet, legs, and powerful quarters enable them to maintain balanced traction. They also carry people, thresh grain, press sugar cane, haul logs, and more. Buffaloes have an advantage over other draught animals with large hooves in wet or muddy areas. Their legs can withstand wet conditions better than cattle.

References

N. Mingala, M. A. Villanueva and L. C. Cruz, "River and Swamp Buffaloes: History, Distribution and their Characteristics," in The Buffalo (Bubalus bubalis) - Production and Research, Bentham Science Publishers, 2017, pp. 3-31.
Zhang, L. Colli and J. Barker, "Asian water buffalo: domestication, history and genetics," Animal Genetics, vol. 51, 2020.
Naveena, M. Kiran, R. Banerjee and M. Muthukumar, "Water buffalo," Elsevier Ltd., 2022, pp. 1-20.
FAO, "FAOSTAT 2018," 2018. [Online].
Mathivanan, "Breeds of Buffaloes," TNAU, 2014. [Online]. Available: https://agritech.tnau.ac.in/animal_husbandry/animhus_buffalo%20breeds.html.
Z. M. Minervino, D. Vecchio and B. Antonio, "Bubalus bubalis: A Short Story," Frontiers in Veterinary Science, vol. 7, 2020.
WR, "The water buffalo: a review," British Veterinary Journal, vol. 137, p. 8–10, 1981.
Wanapat, K. Sommart, C. Wachirapakorn, S. Uriyapongson and C. Wattanachant, "Recent advances in swamp buffalo nutrition and feeding," in 1st Asian Buffalo Association Congress, Khon Kaen University, Khon Kaen, 1994.
Bilal, M. Suleman and A. Raziq, "Buffalo: black gold of Pakistan," Livestock research for rural development, vol. 18, no. 9, pp. 140-151, 2006.
Singh, "The Beef About Buff," Outlook (India), 2016. [Online]. Available: https://www.outlookindia.com/magazine/story/the-beef-about-buff/297825/.
Hill, Cattle and Buffalo Meat Production in the Tropics, Longman Scientific & Technical, 1988.

Rearing and housing system

Buffalo are reared worldwide in five systems — extensive or free-range (bathan), household or backyard, semi-bathan, semi-intensive, and intensive rearing system.

Extensive or free-range rearing system (Bathan)

An extensive or free-range rearing system is buffalo's most common rearing system. Most of the world's buffaloes are found in Asia; an extensive rearing system is the most economical here. In this system, no house is provided for the buffaloes, and kept in the open field throughout the year. In Bangladesh, this system is renamed the ‘Bathan' system. In this system, 51-200 buffaloes are reared in an open space, specifically coastal, river basin, and mostly island areas in Bangladesh. Farmers also provide a fenced area for buffalo calves called ‘Killa (raised land)’ to protect them from predators at night, flood, etc.

Advantage:

Less expensive as no feed supplement is given to the buffaloes.
No shelter is needed.
They can graze in natural pasture land and express natural behavior like wallowing in canals or rivers.

Disadvantages:

Shortage of feed during the dry season and fresh drinking water.
Close monitoring is not possible.
Lack of veterinary and transport facilities.
Loss of animals due to natural calamities as no shelter for the buffaloes is provided.
Clean milk production is not possible.

Household or backyard rearing system

In this system, buffaloes are kept in an open yard near the farmer’s house at milking time, and then they are allowed to graze the whole day. Farmers separate the calf from the dam in the evening and feed the buffaloes with straw and sometimes a little concentrate supplement. Sometimes the buffaloes are kept in a house made of brick or tin.

Advantages:

Close monitoring is possible in this system.
Easy to provide veterinary service.
Hygienic milking is possible.

Disadvantage:

Owners need to be concerned about providing feed
Owners should spend money on shed construction.

Semi-bathan system

It is a combination of Bathan and a backyard-rearing system. In this system, buffaloes can graze freely in an open field or island for most of the year, as in the Bathan system. But they are brought back to the mainland near the farmer’s house during the rainy season.

Advantages:

Less expensive as no feed supplement is given to the buffaloes.
No shelter is needed.
They can graze in natural pasture land and express natural behavior like wallowing in canals or rivers.

Disadvantages:

Shortage of feed during the dry season and fresh drinking water.
Close monitoring is not possible.
Lack of veterinary and transport facilities during staying on the island.
Higher transportation cost of animals to bring back to the mainland.
Loss of animals due to natural calamities as no shelter for the buffaloes is provided.
Clean milk production is challenging.

Intensive rearing system

Farmers keep buffaloes in the shed all day in this farming system. Feed and water are supplied to the buffaloes manually, and farmers provide a shower in the farmyard for wallowing.

Advantages:

Intense monitoring of animals is possible.
Hygienic milk production is possible.
Proper recording can be maintained.
Accessible treatment of diseased animals.

Disadvantages:

Animals cannot express their natural behavior, like wallowing.
A higher density of animals may lead to disease outbreaks.

Semi-intensive system

In this system, buffaloes are allowed to graze in the morning and are provided with concentrate and roughage in the shed. Wallowing can be done using a sprayer inside or in a pond outside the shed.

Advantages:

Close and intense monitoring of animals is possible.
Hygienic milk production is possible.
Proper recording can be maintained.
Accessible treatment of diseased animals.
Buffaloes can exercise in an open field in this system.

Disadvantages:

A higher density of animals may lead to disease outbreaks.

Construction of a buffalo shed

A shed protects the buffaloes from harsh weather conditions, like rain, extreme heat, cold, etc., and wild predators. It also helps in better care and supervision of the animals. It should be comfortable, durable, and have a proper drainage system and facilities for clean milk production.

A shed for buffalo should meet some requirements that will ease the management. They are:

Wall: The wall should be as tall as five ft. from three shade sides. The remaining side would consist of a manger.
Manger: The manger should allow feeding the animals quickly and adequately. There should be a wide (5 ft. in length), non-slippery, easy-to-clean floor with a little slope. The space requirements for manger construction are given below.

Space requirements for the manger of the buffalo shade

Category	Adult	Calves
Inner wall from the ground	50cm	20-25 cm
Depth	≤40cm	≤20cm
Area needed	2.5-3 ft./animal	1.5 ft./animal

Space requirements

Space (floor) requirements for buffalo

Animal type	Floor space/animal (sq. ft.)		Manger length (ft.)
Animal type	Covered	Open	Manger length (ft.)
Adult buffalo	30-40	80-100	2.5
Pregnant Buffalo	100-120	180-200	2.5
Bull	120-140	200-250	2.5
Buffalo calf	20-25	50-60	1.5

Wallowing

Buffaloes have a few sweat glands and dark skin, making maintaining their body temperature in hot and humid environments difficult. To solve this problem, they wallow.

Wallowing means rolling or floundering in mud or water. To the buffalo, it is a defense mechanism against higher temperature and humidity and against ectoparasites (ticks, flies, mosquitoes, lice, etc.). Buffalo wallows for hours as this process is complementary to the sweating process of other species of animals. The buffaloes of the Indian subcontinent prefer wallowing in the clear water of streams and pools huddling close together. On the other hand, the buffaloes of China and Southeast Asia prefer wallowing in muddy water. Due to high humidity, buffalo even wallows or rolls in mud during the cold season.

Figure STYLEREF 1 \s 2. SEQ Figure \* ARABIC \s 1 5: Wallowing behaviour

Wallowing is an acquired behavior. Buffalo behaves more like cattle if no water or mud is available. They will seek cover and graze more in the cooler hours and less in the warmer hours. In the intensive rearing system, farmers can provide shade with a fan, offer abundant water to the animal, and feeding-energy diets and by-pass protein (such as fish meal). The farmers can also allow night grazing for the buffaloes, which will allow increased heat loss from the animal. Farmers can also use sprinklers to spray water on the buffaloes, which can help maintain the body temperature of the buffaloes.

Feeding and nutrition

Feeding systems

Buffaloes have higher digestive efficiency regarding crude fiber and protein than other domestic ruminants (cattle, goats, and sheep). They are better converters of poor-quality feed and forage. They have a rumen with high bacterial activity, a low outflow rate, and a large volume.

Figure STYLEREF 1 \s 2. SEQ Figure \* ARABIC \s 1 6: A farmer offering straw to a buffalo in a household feeding system

Buffaloes can utilize poor-quality roughage, such as rice straw, wheat straw, and grass (low-quality grass, like the remaining part after grazing of cows), and produce more live body weight and milk per day compared to cattle and other ruminants. For this reason, farmers in the tropical region often adopt an extensive feeding system for their buffaloes. A saying is, "If you have ¾ cows, you can rear one buffalo with the left-over feed."

There are four feeding systems in buffalo. Those are:

Free-range or Bathan system: Buffaloes graze freely in an open field throughout the year. No additional feed supplements are given to the animals in this system.
Household system: Household-reared buffaloes are allowed to graze on natural pasture in the field or roadside grass for the whole day, tethered in a poll. Straw is given to the buffaloes in the evening after bringing them back home. The pregnant and lactating buffaloes are given a small amount of concentrate.
Semi-bathan system: Buffaloes in this system can graze freely in an open field for almost a year (7-8 months). But during the rainy season, they are brought back to the homestead. The farmers provide them with grass and crop residues during this period.
Intensive system: In this system, buffaloes remain inside the farm, and grazing is not practiced. Roughages and concentrates are provided to the animals as needed.
Semi-intensive system: Buffaloes are given roughage and concentrate when staying in the shed. After milking in the morning, they can graze in an open area.

Feed ingredients/composition

Buffaloes are fed daily roughages, such as rice straw, wheat straw, maize stalk, and green grass because they are widely available and cheap. Some local feed resources, i.e., such as sugarcane tops, cassava root, Leucaena leaves, and potato vines, are used as unconventional feed sources.

Ruminants have relied on microbial fermentation to meet their daily nutritional requirements. Like other ruminants, buffaloes also utilize their ruminal microbes to digest the feed. Unlike other ruminants, buffalo's rumen has higher digestibility of nitrogen and carbohydrates, promoting the growth and synthesis of rumen bacteria, even when fed diets with low protein content. They also need less supplementary nitrogen in their diet than cattle, as they can efficiently recycle nitrogen from their diet by shunting nitrogen to a higher concentration in the kidneys and extracting a more considerable amount for recycling.

Buffaloes need concentrate (i.e., wheat bran, rice bran, rice police) less than 30% of their diet. A rule of thumb for growing buffalo (around 250 kg) states that 2 kg grain, 750gm protein per animal per day with enough roughage can be followed to gain proper growth.

Breeding

Reproductive characteristics of buffalo

Female Buffalo:

Criteria	Value
Estrous cycle type	Seasonal polyestrous short-day
Age of puberty	26-36 months (average 24 months of age)
Gestation period	300-320 days
Dry period	60-120 days
Lactation period	270 days (average)
Estrous cycle	16-33 days (maximum 21-24 days)
Duration of estrus period	5-27 hours (average 20 hours)
Ovulation	Between 24-48 hours after the onset of estrus or 6-21 hours after the end of the estrus
Anestrous period	25-48 days in the autumn 58-186 days in the summer
Calving season	All the year round, but more tendency in the rainy season (July to September), followed by the winter season (October to January).

Miscellaneous:

Signs of estrus:
The uterus is toned and turgid.
- Discharge of transparent and sticky mucus from the vulva, which changes from cloudy to opaque later. (For more, see 'Heat detection of buffaloes)
- Buffalo cows have a hyper-pigmented vulva and lighter, more rigid, and tortuous reproductive tract with more muscle tone than cows.
Female buffaloes can have two calves in 3 years.

Buffalo Bull:

Age of puberty: 22-24 months of age.
The testicles, accessory sex glands, and penis of male buffaloes are smaller than those of cattle bulls.
The sexual behavior of the buffalo bull is similar to that of cattle bulls but less intense.
Male shows flehmen behavior. This behavior happens when an adult male nears a female in heat. The bull generally opens his mouth, draws the upper lip extending his neck upwards. It appears that the bull tries to inhale or suck both through the nostrils and the mouth. The bull also licks the vulva of the female sometime in this period. Simultaneously the bull also makes some soft penile movements, protruding his penis a few centimeters from the non-motile prepuce.

Heat detection of buffalo

Heat or estrus detection is crucial for every farmer as it is crucial to inseminate buffalo cows with semen naturally or artificially. But the buffaloes show poor expression of estrus, or silent estrus, lack of vocalization, and irregular estrous. They also peak their sexual activity during nighttime due to the adverse effects of the hot environment on their heat expression.

Buffaloes in countries away from the equator line show seasonal breeding characteristics. Most of the buffaloes come to estrus during the winter season more than the summer season. In the summer, estrus often passes unnoticed by the farmers, and the signs of heat may be faint or absent. Estrus is mainly expressed during late-night or early mornings in tropical and subtropical areas.

Farmers often detect heat in buffaloes by observing some signs. These signs are the release of vaginal discharge, bellowing, edema of the vulva, frequent urination, restlessness, and temporary teat engorgement. In a large herd, farmers use teaser bulls to detect heat in the female. Teaser bull is simply a vasectomized/caudal epididymectomy male animal. The courtship and the allowance of mounting by teaser bull indicate the start and the end of the estrus period, respectively.

Non-visual methods of estrus detection involve laboratory examination of vaginal mucous, determination of changes in hormonal levels in plasma and milk, monitoring body temperature, and measurement of variation in physical activity.

Clinical methods of estrus detection are trans-rectal palpation and trans-rectal ultrasonography. Cervical mucous can also be examined in a laboratory to detect estrus. These methods are helpful to farmers in monitoring each buffalo, but it is impractical for herds as they require daily interventions.

There are some new chemical methods for detecting heat in female buffaloes, such as salivary electrolytes and chemical pheromones in the vaginal mucous, urine, or feces. These methods have shown promise for estrus detection in buffalo and are likely to be available in an easy-to-use method soon.

Breeding system of buffalo

The buffalo is polyestrous, and breeding can occur throughout the year, but rainfall, feed supply, ambient temperature, and photoperiod influence the annual calving pattern.

Generally, buffalo's breeding system is of two types — Natural and Artificial.

Natural breeding system: A natural breeding system is when the bull is used for natural service. It is the common practice of buffalo breeding all over the world. Male buffalo plays courtship with the female (during the heat period), mounts the female, and ejaculates his semen inside the female genital tract by thrusting. Farmers use their male buffalo in this system.
Artificial breeding systems: It means mating the male and female reproductive cells with the help of humans and technology.

Artificial breeding systems

There are several methods of artificial breeding systems for animals. But very few methods are used in buffalo. Some of these are:

Artificial insemination (A.I.): It means the deposition of a male's sperm in the female genital tract (specifically the uterus) at a specific moment by using instruments (A.I. gun, vaginal speculum, etc.). This method is commonly used in India, Sri Lanka, China, Malaysia, the Philippines, and other countries. It is a proven technology for the speedy progress of progeny improvement programs. This method is widely used to perform selective breeding, improve an animal's genetics, and overcome the infertility problem in the male.

There are several advantages of A.I. in Buffalo:

No need of breeding bulls in the herd, which is safe for the farmers. Moreover, the cost of maintenance of the bull is minimized.
It helps prevent several genital diseases (brucellosis, trypanosomiasis, vibriosis, etc.) and overall biosecurity of the farm.
Early detection of inferior males and better breeding policy-making can be ensured as regular examination of semen helps to examine the male's fertility status.
Progeny testing at an early age can be done.
The semen of the superior male can be used for a long time after the male is dead.
It ensures the mating of buffaloes with differences in size without physical injury to either animal. It also provides choosing different bulls to match the individual females' size and genetic materials.
It promotes the maintenance of proper breeding and calving records.
Old, heavy, and injured sires with superior traits can be utilized to produce progeny.
Embryo transfer technology (ETT): A superior female is selected as a donor in this technology. She will donate her eggs or embryos to a recipient or surrogate mother. The embryo is transferred, obtained after in-vitro fertilization and in-vitro maturation, into the recipient's uterus to develop a full-term live fetus.

This technology increases the reproduction rate of superior female animals. It also reduces the risk of transmission of infectious diseases in the herd through natural mating.

The ova from the donor can be cryopreserved for further use or transport to rural and urban areas. This process allows the improvement of a breed nationwide. But this technology is comparatively new and costly, and buffaloes were recently inserted into this research program. Maybe this method will soon be adopted as a convenient method for breeding buffalo.

Milking hygiene

Maintaining hygiene during milking is essential for obtaining safe milk and good udder health. Usually, farmers perform milking in buffaloes one time (morning) a day in Bangladesh as milk production is less. It is considered an ideal milking interval in the country for buffaloes.

Bacteria may enter the milk in the following ways:

Animal sources

Animal feces or urine coming into direct contact with the milk
Infection of the udder (mastitis)
Cow diseases (for example, bovine tuberculosis)
Bacteria that live on the skin of animals

Environmental sources

Environment (for example, feces, dirt, and processing equipment)
Insects, rodents, and other animal vectors
Unsanitary conditions in milk processing plant
Contaminated dairy utensil

Human sources

Contamination from dairy workers, such as contact with dirty hands, clothing, or boots

Pre-requisites of hygiene milking

Grooming, washing, and drying

While the animals should be carefully groomed and washed, the udder and teats should not be submerged in water. If there are any long hairs in the tail or on the udder, they should be removed. The udder and teat should be dried as soon as possible after being washed, paying attention to the udder and teats. A towel can be used for drying. It is also advised to use one towel per buffalo on the farm. This practice will allow less cross-contamination between animals during milking.

Preparation

The animals should be milked simultaneously and in the same procedure daily. The same milker should be milking the same animal(s). Sudden changes in the milking time, procedure, and milker may act as a risk of mastitis.

The milker should wear a clean uniform and disposable gloves, and their nails should be trimmed. If not changing gloves is impossible, the milker can use alcohol spray each time after milking. The milker should not be allowed to abuse the animals physically, i.e., beating and making the animal uncomfortable.

Feeding during milking

Cows should be given concentrates during milking to enhance milking. Cows should be given green grass after milking to make them stand for at least 30 minutes, but ideally 1 hour. This process lets the teat ends seal, reducing the risk of mastitis.

Milk container (can)

Milk should be temporarily stored in a standard milk container as cool as possible that has been well sanitized. To make cleaning easier, aluminum or stainless-steel milk cans should be used instead of plastic ones. Immediately following milking and emptying, the milk container should be cleaned. It has to be washed in cold water, scrubbed with a brush in hot water with detergent, rinsed in cold water once more, and then kept in a well-ventilated space to dry naturally. The can should be left upside-down on a clean surface until the subsequent usage to keep dirt from getting inside.

The procedure of hygienic milking

Preparation of milker:

Milker should wear a clean uniform and a pair of fresh, sterile gloves after washing and drying of hands. Disposable gloves are the most effective in hygienic milking procedures, but the farmer can also use non-disposable gloves. In that case, the gloves should wash thoroughly after the milking, and can only use one pair of gloves per animal.

Fore stripping:

Before actual milking, it is best to examine the first few squirts of milk to detect any changes in the color and consistency of the milk. Also, dirt present in the teat canal will be removed during the fore stripping.

Cleaning of udder and teats:

The udder and teats should be cleaned with a damp cloth (one for each animal) to wash any impurities in the hindquarter.

Pre-dipping:

The teats are pre-dipped with a low concentration of antiseptic (e.g., 0.1% iodophor, low concentration of chlorhexidine, lactic acid, and chlorine dioxide) to prevent contamination.

Drying:

After pre-dipping, the teats should be wiped with a clean towel, cloth, or paper towel. It is an excellent practice if separate towels or cloth are used for individual animals. After wiping the teats, the towel or cloth should be washed at a high temperature.

Stimulation:

The udder should be stimulated by touching or massaging to arouse the milk let-down process. Washing and drying are also another way of stimulation in the milking process. The presence of calf and suckling also stimulate milking.

Milking:

Milking is performed in two ways – hand milking and machine milking. Both are described in the "Milking system" section.

Post-dipping:

After milking, the teats are again dipped in an antiseptic solution. It is called post-dipping. It helps clean and disinfect the teat surface by eliminating the microorganisms on the skin surface. The solution should not be wiped or dried off manually.

Milking system

There are two systems of milking in buffalo all over the world.

Hand milking
Machine milking

Hand milking

Milking is performed by the hand of the milker. The milking starts pairwise from the left/right side or hind/front teat. Teats should be squeezed rather than pulled to avoid laceration or tissue damage.

Figure 2.9: Hand milking. Picture credit: collected from web

Hand milking is four types –

a. Full hand milking: It is the standard method of hand milking where the teats are pressed between the palm and three lower fingers of the hand while keeping the teat base closed between the index finger and thumb.

Full hand milking. Picture credit: collected from web

b. Two-finger method: It is also called stripping and is used for cows with very short teats. For instance, cows after 1st parturition usually have small-size teats, making this method more suitable.

Two-finger method. Picture credit: collected from web

c. Knuckling/Fisting: Bend the thumb against the teat and hold the teat with the other four fingers. Some farmers practice this method though it may cause teat injury

d. Full hand milking followed by stripping: The teat is grasped with all the fingers and pressed against the palm, followed by stripping.

Machine milking

Most milking in developed countries is done using milking machines. Teat cups are attached to the cow's teats, and then the cups alternate between vacuum and normal air pressure to extract the milk. The milk is filtered and cooled before being added to a large bulk tank for storage.

Figure 2.12: Machine milking. Picture credit: collected from web

Machine milking requires less labor, allowing less contamination in the milk if performed correctly. In Bangladesh, machine milking is not practiced in buffaloes. There are three types of machine milking used in buffalo farms. They are –

a. Portable milking machine: A portable milking device uses a motor with batteries that can be easily transported inside the farm.
b. Barn milking machine: The machine gets power through the farm's electricity. It is transportable in the cable range.
c. Milking parlor: Milking parlor is applicable in large-scale organized commercial farms. The animals pass through the milking parlor and get milked by the milking machines automatically.

Figure STYLEREF 1 \s 2. SEQ Figure \* ARABIC \s 1 13: Portable milking

References

S. P. Sahu, "housing and rearing systems for cattle and buffaloes," 2017. [Online]. Available: https://basu.org.in/wp-content/uploads/2020/10/LPM-601_Housing_and_Rearing_Systems_for_cattle_and_buffaloes.pdf
ICAR, "Buffalo Housing," Indian Council of Agricultural Research, 2019. [Online]. Available: https://buffalopedianew.cirb.res.in/buffalo-housing/
F. Napolitano, C. Pacelli, F. Grasso, A. Braghieri and G. De Rosa, "The behaviour and welfare of buffaloes (Bubalus bubalis) in modern dairy enterprises," Animal, vol. 7, no. 10, pp. 1704-1713, 2013.
C. Bhakat, "Wallowing in Buffalo and summer management," 2012. [Online]. Available: https://osf.io/preprints/indiarxiv/juctz/download
B. Lemcke, The Australian Water Buffalo Manual, Department of Primary Industry and Resources, 2017.
G. N. Purohit and T. K. S. Rao, "Estrus Detection in Buffaloes," Bubaline Theriogenology, 2018.

M. A. Zava and M. Sansinena, "Buffalo Dairy Production: A Review," in The Buffalo (Bubalus bubalis) - Production and Research, Bentham Science Publishers, 2017, pp. 225-261.

3.1 Anatomy and Physiology of the Udder

Buffaloes have four mammary glands with teats: two forequarters and two hindquarters. Every teat has its orifice. The organ is located in the animal's ventral inguinal area. Milk production is the organ's primary purpose.

Figure 3.1: Buffalo udder. Picture credit: collected from web

3.1.1 Anatomy of the Udder

3.1.1.1 External anatomy

Except for the teats, the udder is hairy. Bowl-shaped udders are the most prevalent, but rounded, pendulous, and goaty udders also exist. The teats are long, black, and conical/cylindrical/bottle/funnel-shaped. Milk production depends on udder shape. The highest average daily milk output was in Murrah buffaloes with bowl-shaped udders, followed by pendulous, globular, and goaty udders. The teats are 5.9-6.9 cm long. The glands lie in two rows on either side of the body's median line. Each quarter has two typical functioning glands. A longitudinal intermammary groove or sulcus intermammaricus divides the right and left parts of the udder. There is no apparent septum between the front and rear quarters. Typically, one teat empties one gland. The teat wall has no sweat or sebaceous glands.

Figure 3.2: Different udder shapes (Photo credit: Raju, 2008)

The udder is well-attached to the body and muscles. Elastic fibrous tissue makes up the middle ligaments, while less elastic connective tissue makes up the lateral ligaments.

Figure 3.3: External features of a bovine udder. Picture credit: collected from web

3.1.2 Internal anatomy

The mammary gland has secreting and connective tissue. The number of secreting cells or tissue limits the udder's milk production. The secretory cells, which form a single layer on a basal membrane in alveoli, synthesize milk. Each alveolus has a diameter of 50-250 mm.

Figure 3.4: Internal anatomy of a bovine udder. Picture credit: collected from web

Several alveoli create a lobule. The alveoli, milk ducts, udder, and teat cistern contain milk continually synthesized in the alveolar area between calf milkings.

Teats have a canal and a cistern (Sinus papillaris). Where the teat cistern and teat canal meet, 6-10 longitudinal folds form Fürstenberg's rosette, which fights mastitis locally. Longitudinal and circular smooth muscle bundles border the teat canal. The smooth muscles close the teat canal between milking or suckling by the calf. Keratin or keratin-like proteins in the teat canal block harmful bacteria between calf milkings.

3.1.3 Physiology

The mammary gland is dependent on the hormone cycle. Before puberty, the growth of simple ducts occurs only in pads of fat, whereas after puberty, there is growth of ducts only at a greater rate. In pregnancy, secretory alveoli are 'budded on' to the enlarged duct system under the influence of the corpus luteum. During pregnancy, alveoli mature, and milk precursors are accumulated.

After parturition, the anterior pituitary gland controls milk formation and secretion maintenance. The release of milk at suckling or mechanical milking is mediated via oxytocin from the posterior pituitary.

Milk is produced in the alveoli by alveolar cells synthesizing fat, casein, and lactose. Selective permeability of alveolar cells to blood constituents, i.e., calcium, potassium, and phosphates, occurs. During involution, when the udder goes from lactating to dry, the alveoli stop secretion and shrink with the desquamation of cells. Hence, there is an increase in cell number in the milk at the end of lactation.

3.2 Mastitis

Mastitis is a widespread disease of the udder in buffalo and one of the significant problems for the dairy industry since it negatively impacts milk quality and quantity, thus causing heavy economic loss. Farmers' and veterinarians' non-judicious use of antimicrobials to treat diseases may develop antimicrobial resistance and antimicrobial residues in milk. It may compromise public health safety in the consumption of milk.

3.2.1 Types

Mastitis can be classified in a variety of ways; however, a classification based on symptoms divides it into two broad groups:

a. Clinical mastitis: Clinical mastitis (CM) is defined by visible changes to milk, the udder, and the body system. CM is further classified as mild, moderate, and severe based on the severity of the inflammation –

b. Subclinical mastitis: In subclinical mastitis (SCM), no apparent abnormalities in the udder or milk, yet milk output diminishes as the somatic cell count rises (SCC). It is 15-40 times more prevalent than CM in Buffalo.

A specific type of grade-3 mastitis is gangrenous mastitis. This type of mastitis causes udder congestion and edema. The damaged area is bluish and spreads to the udder’s ventral side and other teat components. Palpating the udder feels cold. Animals can die from improper care. Even with quick treatment, the affected quarter may be damaged, and the gangrenous area may be sloughed.

Figure 3.7: Enlarged left hind teat (white arrow). Credit: Sangwan, et al. 2018

Mastitis can also be categorized based on duration, with acute cases showing a fast onset and chronic issues offering a longer course. Acute or chronic mastitis can occur in both clinical and subclinical cases. Mastitis can also be classified as infectious, caused by microorganisms, or non-infectious, induced by trauma.

3.2.2 Causal agents

Pathogens causing mastitis in water buffalo are mainly bacteria but occasionally can also be fungi and algae. The most common mastitis-causing bacteria of mastitis include Staphylococcus (S.) aureus, non-aureus staphylococci (NAS), Corynebacterium bovis, Streptococcus (Strep.) agalactiae, Strep. uberis, Strep. dysgalactiae, Escherichia coli, Klebsiella spp, and Pseudomonas spp are some of the most prevalent mastitis-causing bacteria in water buffalo.

Another way to classify mastitis-causing pathogens is by their route of transmission:

Contagious or cow-bound pathogens are transmitted mainly from cow to cow (or udder to udder) and during milking. Infectious bacteria include agalactiae, S. aureus, and Mycoplasma bovis. Contagious pathogens are best controlled by good biosecurity and optimal milking routines.
Environmental pathogens are mainly transmitted from the environment and mainly between milkings. Examples of environmental bacteria are coli, Klebsiella, and Strep. uberis. Environmental bacteria are best controlled by good hygiene and by strengthening the cow's immunity.

3.2.2.1 Uncommon Mastitis Pathogens

Some uncommon pathogens can cause mastitis in buffalo.

Type of pathogen	Pathogen name
Bacteria	Enterococcus faecalis, Serratia spp., Leptospira spp., Nocardia spp., Pasteurella multocida, Mannheimia haemolytica, Listeria spp., Campylobacter jejuni, Bacillus cereus
Fungi	Trichosporon spp., Aspergillus fumigatus, Candida spp.
Yeast	Cryptococcus neoformans and Saccharomyces spp.
Algae	Prototheca spp.

3.2.3 Risk factors

Mastitis is a multifactorial disease, and several risk factors are involved in the epidemiology of the disease. Some risk factors for mastitis and intramammary infection (IMI) are.

Animal-related risk factors
- Age and parity: The prevalence of infected quarters increases with age, peaking at seven years. In addition, as the parity increase, so do the incidence of mastitis.
- Stage of lactation: Most new infections occur during the early dry period and the first two months of lactation, particularly with environmental pathogens.
- Breed: It has been discovered that pure-breed buffalo have clinical and sub-clinical mastitis 14% more frequently than crossbred buffaloes.
- Udder and teat conformation: Mastitis is more prevalent in animals with bowl or circular udders than the cup, pendulous, or teat/udder lesions. Larger teat apex/base diameters and shorter teat lengths help microorganisms to penetrate the teat orifice and infect breast tissue faster, causing mastitis.
- Milk yield: Higher milk-yielding animals are more susceptible to mastitis than low-yielding animals.
- Suckling: Calf sucking affects the cow's udder health. It was found that a longer duration of calf suckling decreases the risk of mastitis. Boden and Leaver (1994) showed a substantial drop in milk SCC in cows milked once daily (morning) and suckled twice daily by their calves.
- Previous history of mastitis: Buffalo cows with prior history of clinical mastitis are more prone (almost twice as susceptible) to the disease than cows without any previous history of mastitis.
- Body condition score: Cows with poor body condition score (BCS) are more susceptible to mastitis than cows with medium and good BCS.
- Other concurrent diseases: When the buffalo's immune system is already compromised due to concurrent diseases, mastitis is more likely to develop. Retained placentas, teat injuries, and ulcers may also increase the risk of mastitis. The severity of sole ulceration, e.g., more than one finger, has been linked to a three-fold increased incidence of aureus infections in the first lactation. Sore feet may also cause teat lesions due to trouble standing.
Environment and management risk factors
- Bedding material: Mastitis is less common on sand flooring than concrete and more common when soil is used as bedding material.
- Milking method: The hand milking method, e.g., the knuckling method, increases the prevalence of clinical and subclinical mastitis compared to full-hand. Compared to hand milking, milking machines reduce mastitis in water buffalo.
- Milking routine:
  - Washing milkers' hands before milking results in less mastitis than farms that don't wash their hands before milking.
  - Farms that did not use a drying towel for each cow's udder had substantially higher rates of mastitis than those that did.
  - Using teat dips before and after milking reduces mastitis.
- Feeding system: The dairy buffalo needs more energy and nutrients to produce colostrum and milk during lactation. The buffalo has a negative energy balance when feed intake doesn't fulfill lactation demands. Notably, negative energy balance causes diet deficiencies in trace elements (selenium, iron, copper, zinc), amino acids, and vitamins (A, C, E) that cause immunosuppression during lactation and increase infection risk. Thus, vitamin E and zinc supplementation during the transition are essential to prevent mastitis and increase milk.
- Herd size: The buffalo herd size may positively correlate to mastitis because infectious mastitis may be harder to control in a herd with more IMIs and animal-to-animal contacts.
- Farm biosecurity:
  - External biosecurity: External biosecurity includes all measures required to prevent both the entry of infectious pathogens into farms and the dissemination of infectious agents from farms.
    - Keeping newly purchased animals in the quarantine shed for 3-4 weeks will reduce the risk of introducing diseases (even subclinical infections like SCM) in the herd.
    - Including antiseptics in the milking routine and at the shed's entrance prevents microbial contamination in the herd.
  - Internal biosecurity: Internal biosecurity includes those measures highly connected with farm management and daily practices on the farm.
    - Keeping the milking area and other areas clean and free of manure and other debris can help prevent the spread of pathogens.
    - Vaccinating buffalo against diseases that can cause mastitis, such as agalactiae, can help to reduce the risk of infection.
    - Keeping records of where animals have been and who they have come into contact with can help to identify and track down the source of an outbreak of mastitis. Isolating the mastitis-affected cows and grouping them during milking is necessary to control the spread of infection.

Viability of pathogens: Each pathogen can live in the buffalo's local environment (resistance to environmental conditions like washing and disinfection). They're opportunistic pathogens. They can enter the teat during milking or when the cow's immune system declines, causing clinical mastitis

3.2.4 Diagnosis

Clinical symptoms, somatic cell count (SCC), white blood cells in milk, and bacteriological investigations are utilized to diagnose mastitis and IMI. Bulk milk SCC is used to evaluate udder health in herds.

Physical appearance: Clinical mastitis is diagnosed by abnormal milk, which may be

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 8: Normal milk (A, B) and clots in mastitis milk (C, D) (Crefit: From the web)

Strip cup test: The operators visually evaluate the milk for gross abnormalities by squirting a few streaks of milk on a strip cup, where the common abnormalities are the presence of blood, flakes, clots, and wateriness suggestive of mastitis.

Figure 3.9: Strip cup test (Credit: From the web)
Direct SCC: The direct microscopic somatic cell count (DMSCC), bulk milk somatic cell count (BMSCC), and individual cow somatic cell count (SCC) are used to interpret milk SCC. Buffalo quarter milk with < 200,000 cells/mL SCCs considered healthy. The presence of SCM quarter is indicated by an SCC of 200,000 cells/ml.
California mastitis test (CMT): CMT is a fast, inexpensive cow-side screening test. It is done by combining CMT reagent with an equal volume of milk. The reagent creates a gel to measure SCC indirectly [17]. As shown in table-1, CMT scores and average SCC scores are correlated.

Figure 3.10: California Mastitis Test. Credit: Raúl Mazo Velásquez

Score	Observation	Somatic cell range (cells/ml)	Picture
1	No thickening of the milk (Negative)	˂200,000
2	Milk thickens slightly. Trace reaction disappears with paddle rotation. (Trace)	150,000-500,000
3	Milk thickens without gel formation. Thickening may disappear after 20 seconds of CMT paddle rotation.	400,000-1,500,000
4	Milk gels immediately. The bottom of the cup is shown as the milk swirls toward the middle. The milk settles and covers the cup's base after the action stops.	800,000-5,000,000
5	A gel forms and the surface of the milk rises like a cooked egg. Central peak persists after CMT paddle rotation stops.	˃5,000,000

Table 3.1: Relation between CMT scores and average SCC

Direct SCC: The direct microscopic somatic cell count (DMSCC), bulk milk somatic cell count (BMSCC), and individual cow somatic cell count (SCC) are used to interpret milk SCC. Buffalo quarter milk with < 200,000 cells/mL SCCs considered healthy. The presence of SCM quarter is indicated by an SCC of 200,000 cells/ml.

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 11: Somatic cell counter
- Bacteriological culture: A sterile milk sample collection is necessary to perform a reliable bacteriological examination of the sample. Individual cows or bulk milk should be submitted for the presence of an IMI either as a routine practice before drying off or to identify the etiology of SCM or CM cases. An IMI caused by aureus or S. agalactiae is a reliable parameter for the possibility of contagious bacterial transmission in the herd. The standard procedure of sterile milk sample collection is given below.
Sterile milk collection:

Milk samples from individual quarters should be taken from the closest teat first. Wash the teat with clean water and alcohol. Don't put the cap down or touch the inside during sample collection. Milk collection tubes should be held at a 45° angle to prevent the teat's dirt from entering the collection tube. There should be a distance between the teat and the tube so that the teat would not touch inside the tube. Hands and fingers should not touch milk or the tubes' inner wall. Sample tubes should never exceed three-fourths capacity. Finally, transfer these tubes to an ice box.

Figure 3.12: Procedure of sterile milk sample collection
Electrical conductivity test: Checking milk's electrical conductivity can diagnose mastitis. The electrical conductivity of milk can be assessed using a hand-held (portable) electrical conductivity meter (milk checker or digital mastitis detection). Healthy buffalo milk has an EC of 9.17 ± 1.51 Ms/cm.

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 13: pH meter

pH determination test: The normal pH of buffalo milk is 6.70 to 6.81. The milk becomes more alkaline during mastitis and contains more chlorides

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 14: Electrical conductivity meter

3.2.5 Control and prevention

Controlling and preventing mastitis involves reducing teat exposure to infections or increasing dairy animal resistance. A 10-point mastitis control program was created by the NMC (National Mastitis Council) and encouraged specific treatment techniques. This control program includes:

Establishment of goals for udder health
1. Establish realistic targets for the clinical mastitis rate and the average bulk somatic cell count (BMSCC).
Maintenance of a clean, dry, comfortable environment
1. Keep areas clean and dry where cows gather.
2. Make sure the ventilation system is operating correctly.
3. Ensure the animals stand after milking (provide fresh feed and water).
Proper milking procedures
1. Before milking, make sure the teats are clean and dry.
2. If possible, apply pre-milking teat disinfectant (pre-dip), then dry teats with a cleaned and disinfected disposable cloth towel. (See more in "Teat dipping")
3. When milking, use clean gloves to prevent the spread of infectious pathogens.
Proper maintenance and use of milking equipment
1. After each milking, thoroughly clean and sanitize the milking equipment.
Good record keeping
Effective dry cow management
(See in the "Dry cow management in Buffalo" section)
Maintenance of biosecurity for contagious pathogens and marketing of chronically infected cows
1. Obtain aseptically collected milk samples from buffaloes for bacteriological culture before purchasing.
2. Isolate recently purchased buffaloes and milk separately until the absence of intramammary infection is confirmed.
3. Buffaloes with a chronically high SCC score (i.e., SCC >200,000) should be separated and monitored for response to dry treatment or other indicated therapy.
Regular monitoring of udder health status
1. Enroll in an individual Buffalo SCC program or apply other monitoring.
2. Regularly perform milk bacteriological cultures on clinical cases and high SCC buffaloes.
3. Regularly calculate clinical mastitis rates and distributions, giving special attention to infections in heifers.
Periodic review of the mastitis control program
When these strategies are correctly implemented, the prevalence of prevalent infectious mastitis pathogens is dramatically reduced.

3.2.5.1 Dry cow management

What is the dry period?

The dry period of a dairy animal's lactation cycle is crucial. Buffaloes are not milked during this period, generally about 155-220 days before calving. The dry phase normalizes mammary gland epithelial involution, boosting milk production during lactation. Eliminating or reducing the dry period diminishes milk production following parturition.

Advantages

The dry time allows lactating animals to recover. It helps a cow give birth and produce high-quality milk in the subsequent lactation season. The dry period is applied to 1) repair the mammary gland, 2) cure existing infections, and 3) prevent future infections.

Drying-off procedure

Reduce concentrate feeding on a tapering basis 2-3 weeks before drying-off
Stop concentrate feeding 1-2 weeks before drying-off
Restrict feed intake if milk yield is above 12 liters per day
Reduce dry matter allowance to maintenance levels three days before planned dry-off
Stop milk secretion quickly and gently. But do not restrict water access
Check the animal visually daily after drying-off
Teat-dip the first days after the last milking.

Management during the dry period

In buffalo cows, the dry season typically lasts four months. Nutrition during this time should meet needs for the final third of pregnancy and maintain weight. An energy level of 0.65 Mcal/kg dry matter and a protein level of 10.5% are suggested. Fresh, high-quality forages, hay, and 15% dry matter should be fed to animals in addition to concentrates. Low fermentation rates define this period's food. Clean and comfortable conditions should be provided for dry cows.

Dry cow therapy with antibiotics

Dry cow therapy eliminates 70%–98% of infections and prevents new ones. Each quarter should receive a long-acting intra-mammary antibiotic after the buffalo's last milking. Udder tubes containing long-acting benzylpenicillin, ampicillin+cloxacillin, ceftiofur, and gentamicin are used to dry off the cow. Selective dry cow therapy is usually advised.

Types of dry cow therapy

Dry cow therapy is classified into two types:

Blanket dry cow treatment (BDCT): All buffaloes are given antibiotics during the dry season in BDCT. For decades, most dairy farms have used this treatment. Since blanket dry cow treatment treats healthy animals, many countries now use selective dry cow treatment.

Selected dry cow therapy (SDCT): SDCT only treats diseased or all of a buffalo's quarters with at least one affected quarter. It reduces antibiotic resistance and is done at the end of lactation.

3.2.5.2 Teat dipping

Teat dipping or disinfection may help minimize exposure of IMI-causing pathogens into the teat canal, thereby reducing occurrences of mastitis in dairy farms. Teat dipping reduces milk SCC. The disinfection performed before milking is known as pre-dipping, and the disinfection performed after milking is known as post-dipping.

Figure 3.16: Using of teat dipper in bovine (Photo credit: From the web)

Pre-dipping

Pre-dipping involves applying a quick-acting disinfectant to the teat skin before milking to reduce mastitis-causing bacteria, especially in the external teat orifice. Before milking, the teats are dipped in a low concentration of antiseptics. Pre-dipping solutions include 0.1 % iodophor, low-concentration chlorhexidine, lactic acid, and chlorine dioxide. The operator should dry off the teats after dipping them to avoid mixing chemicals with the milk.

Pre-dipping is performed in three steps –

Strip: Wipe loose dirt from each teat and pre-strip foremilk
Dip: Pre-dip and wait for at least 15 seconds, preferably 20-30 seconds
Dry: Wipe each teat with a single-use paper towel

Teat-cups containing disinfectants are applied 60-90 seconds after the first touch of each udder.

Post-dipping

Post-dipping or spraying with a germicidal solution after milking reduces new intramammary infections. The mixture has an emollient and an antiseptic solution in it.

There are many post-dipping solutions in several categories. Iodine (69%), chlorhexidine (13%), fatty acid-based (7%), chlorine (2%), and quaternary ammonium (0.6%) were the most common antiseptic post-dipping solutions.

3.2.6 Treatment strategy

The treatment strategy depends on whether the mastitis is clinical or subclinical, as well as the herd's health and history. Collect valuable data before, during, and after treatment, like:

Identification of the animal
Quarters affected
Severity of symptoms
Date of mastitis event
Lactation number
Date of calving
Identification of pathogen(s)
The treatment used previously, including dose, route, and duration

Principles of treatment of mastitis include:

Aseptically treat the animal. Handle the affected udder quarter with disposable gloves.
The drugs should be non-irritant to the udder tissues. Treatment is administered directly in the udder or systematically during systemic infection.
Narrow-spectrum or specific antibiotics should be used in the treatment of the disease.
Before administering the drug, perform a Cultural Sensitivity (CS) test.
Appropriate management of clinical mastitis during lactation
1. Develop and implement a herd clinical mastitis treatment program with the udder health advisory team.
2. Aseptically collect a pre-treatment milk sample for microbiological culture to apply CS test as needed.
3. Use a regulated drug.
4. Don't treat chronic non-responsive infections.

Antibiotics are indicated during the first six hours of infection with supportive treatment [30]. Antibiotic treatment of animals requires a proper diagnosis and bacterial sensitivity estimation. The welfare of animals must be taken into account. The animal's prognosis, general infection level, and potential spread of resistance genes in the herd may make euthanasia a better option than antibiotics.

Selection of antibiotics

A veterinarian must answer four questions before prescribing antibiotics for mastitis:

Is antibiotic therapy indicated?
1. Is antibiotic treatment necessary, or can supportive therapy be used?
2. Several points should be considered while providing antibiotic therapy –

Types of pathogens involved:
- agalactiae responds hugely well to antibiotics
- aureus mastitis has a poor response to antibiotics
- Environmental mastitis has a variable response to antibiotics
Duration of infection
- Acute cases have a better prognosis.
- Chronic aureus mastitis has a shallow cure rate.

Stage of lactation
- Cost-benefit analysis of mastitis treatment depends on the lactation stage.

Age of the animal
- The chance of recovery from mastitis is higher in young animals.

Which route of administration should be used? Intramammary, parenteral, or both?
1. Antibiotics aim to maintain an effective concentration at the infection site.
2. Intramammary antibiotics work well against infections that only affect the milk and milk ducts (like S. agalactiae and C. bovis). Parenteral antibiotics are best for treating mastitis caused by systemic-infecting bacteria like Klebsiella or udder-invasive bacteria like S. aureus and Trueperella pyogenes. According to studies, intramammary and parenteral administration can improve S. aureus treatment efficacy.
Which antibiotic should be administered?
1. Culture and susceptibility testing has been used to determine the antibiotic class for the mastitis pathogen.
2. The immune system eliminates many Gram-negative bacteria, so no antibiotics are needed. The illness should be treated with only supportive care. In life-threatening cases, antibiotic therapy (e. g., tetracycline) is recommended [30].

Specific antibiotics for specific group organisms are listed in the following tables –

Table 3.2: Choice of antibiotic for G +ve bacteria (β-lactam negative)

Gram positive bacteria (β-lactam negative)

Most streptococci, many staphylococci

First choice

Benzyl-penicillin

Parenteral treatment is recommended

Second choice

Antibiotics with supportive treatment

Table 3.3: Choice of antibiotic for G -ve bacteria (E. coli)

Gram-negative bacteria (E. coli)
First choice	No antibiotics; only supportive treatment
Second choice	Antibiotics for Gram-negative bacteria (e. g., fluoroquinolone) in case of a severe and life-threatening case

Table 3.4: Choice of antibiotic for G -ve bacteria (Klebsiella spp.)

*Gram-negative bacteria (Klebsiella* spp.)**
First choice	Antibiotics for Gram-negative bacteria (e. g., fluoroquinolone)	Parenteral treatment is recommended
Second choice	Antibiotics with supportive treatment

Antibiotic treatment is not advocated for mastitis caused by penicillin-resistant S. aureus, E. coli, Mycoplasma bovis, pseudomonas, fungi, and algae. The infected animals are recommended for culling because of these pathogens' persistent infection and contagious nature.

What should be the frequency and duration of treatment?

1. Parenterally administered antimicrobials are administered according to their pharmacokinetics and pharmacodynamics. For example,

Concentration-dependent antimicrobials like fluoroquinolones (ciprofloxacin, marbofloxacin) and aminoglycosides (streptomycin, gentamicin) kill bacteria faster at higher concentrations.
Time-dependent antimicrobials like macrolides (erythromycin, azithromycin), β-lactams (penicillin, cephalosporin), and lincosamides improve efficacy by exceeding the MIC at the site of infection for a longer percentage of the inter-dosing interval.

1. Since intramammary formulations are mostly cleared by milk withdrawal, administration frequency depends on the milking schedule.
2. Effectiveness is increased by prolonging the use of antibiotics in mastitis treatment.

Treatment of subclinical mastitis

Subclinical mastitis during lactation is rarely treated. Subclinical mastitis should be treated during the dry period. The causative organism, animal age, number of intramammary infections, and herd udder health should all be considered during treatment.

3.3 Other Disorders of the Udder and Teats of Buffalo

Disorders in the udder and teats of buffalo can occur at any time and age. They disturb the normal udder health of the animal. Common disorders of udder and teat are briefly described below:

3.3.1 Disorders of udder

a. Udder edema: Udder edema is a periparturient disorder characterized by excessive accumulation of fluids in the intercellular tissue spaces of the mammary gland. Udder edema can be a significant discomfort to the buffalo. It may cause management problems such as difficult milking, increased risk for teat and udder injuries, mastitis, and reduced milk production. The exact reason for this disorder is unknown, but diet, obesity, and lack of exercise may cause the incidence. In addition, excessive salt intake during the dry period may influence the development of the disorder, as increased Na in the blood may result in fluid accumulation in the udder tissues. The udder and adjacent tissues become swollen, and pits-on-pressure are present. The udder may become painful sometimes.

b. Hematoma: Self-induced trauma resulting from awkward efforts to rise or lie down and external trauma from other cows are theorized as the causes of udder hematomas. Hematomas usually appear as soft tissue swellings located cranial to the fore udder or caudodorsal to the hind udder. Severe hematomas can result in anemia if not treated. Hematomas that continue to enlarge should be considered an emergency because of the possibility of excessive blood loss and shock.

c. Hemolactia: Pink- or red-tinged milk is common after calving and can be attributed to the rupture of small mammary blood vessels. Udder swelling from edema or trauma is a potential underlying cause. Several infections, including those caused by some bacteria (Leptospira spp., Brevibacterium erythrogenes, Serratia marcescens, Micrococcus cerasinus, Sarcina rubra, etc.), some viruses and red yeast (Monascus purpureus) may cause systemic infections associated with intravascular hemolysis and capillary damage in the udder. Certain plant toxins in ranunculi, conifers, poplars, alders, etc., may cause capillary damage. Animals affected with diseases characterized by low platelet count may show reddish or pinkish discoloration of milk due to leakage of blood into milk. Bright red milk from a single quarter is a sign of a larger blood vessel hemorrhage or teat trauma, while dark brown milk results from the rupture of a major vein or severe mastitis. Milking should be discontinued until the hemorrhage is controlled.

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 17: Hemolactia

d. Abscess: Udder abscesses may appear anywhere in the mammary tissue or adjacent to the glands. Abscesses can form secondary to mastitis with abscessation, typical of mastitis caused by Trueperella pyogenes. Abscesses appear as firm, warm swellings that may be distinct or indistinct from the gland parenchyma. Palpation of the swelling may be painful to the affected buffalo. Milk from the affected quarter is usually normal, and the abscesses are well encapsulated.

e. Udder fibrosis: Udder fibrosis is one of the significant problems for dairy buffalo. Chronic mastitis acts as a leading cause of udder fibrosis. Persistent inflammation in the udder tissues in chronic mastitis leads to udder fibrosis, then to udder atrophy. The disorder results in reduced or complete loss of milk production. Chances of recovery of chronically affected quarters are very low once atrophy/fibrosis has occurred.

f. Udder cleft dermatitis: Udder cleft dermatitis is a foul-smelling, painful area of moist dermatitis that results from pressure necrosis of the udder skin associated with periparturient udder engorgement and edema. Common locations include the skin reflection between the medial thigh and dorsal attachment of the lateral udder, on the ventral midline immediately adjacent to the median septum of the fore udder, and the median septum of the udder - either between the forequarters or in the fold that is centered between the four quarters. The condition may aggravate mastitis. High parity, high milk yield, and existing mastitis may be risk factors for this condition.

Figure STYLEREF 1 \s 3. SEQ Figure \* ARABIC \s 1 18: Udder cleft dermatitis

g. Udder impetigo: Udder impetigo (udder acne) is a bacterial dermatitis characterized by the development of small pustules on the skin of the udder and teats. Staphylococci usually are the main causal agents for the condition. Lesions of udder impetigo are numerous small, creamish-yellow, and 2-4 mm diameter pustules on the udder.

3.3.2 Disorders of the teats

a. Supernumerary teat: This common congenital anomaly is characterized by more than four teats. Although the extra teat may be attached through the outer attachment, the teats are entirely separated internally. It is possible to get mastitis in the supernumerary teat, which may cause milking difficulties.

Figure 3.19: Supernumerary teat (Photo credit: From the web)

b. Bovine ulcerative mammillitis: Bovine herpesvirus II and IV cause an acute, ulcerative condition of dairy buffalo's teat and udder skin, referred to as Bovine Ulcerative Mammillitis or Herpes Mammillitis. It occurs sporadically or in seasonal outbreaks often associated with cold weather. The characteristic sign is the presence of raw, ulcerated areas in the teat covered with thick brown scabs. It may reduce milk production and increase susceptibility to bacterial mastitis.

Figure 3.20: Bovine ulcerative mammillitis (Photo credit: From the web)

c. Pseudocowpox: Pseudocowpox virus causes the disease in the teat of buffalo. The lesions first appear as tiny, red papules on the udder or teats. Small vesicles or pustules may appear before scabs do, although they may be quickly followed by scabbing. Scabs may be many and easy to remove painlessly. Under the scabs, there is granulation, which causes a raised lesion that heals from the center and leaves a recognizable horseshoe-shaped or circular ring of tiny scabs.

Figure 3.21: Pseudocowpox. Credit: Paul Gibbs

d. Buffalopox: Caused by buffalopox virus in the family Poxviridae. Lesions start as papules, progress to yellow pustules, and then develop a depressed center. It is a zoonotic disease.

Figure 3.22: Buffalopox lesions in buffalo (a–c). Typical pox lesions are seen on the ear (a, b) and teats (c) of buffalo. Credit: Chandrashekhar Raut

e. Warts/bovine papillomatosis: Warts in the udder of buffalo are caused by bovine papillomavirus (BPV) type 1, 5, and 6. Warts occur in two common forms.

Pale, smooth, raised, benign nodules due to BPV5. They may be highly abundant on the teat skin. Some nodules may persist indefinitely without causing problems; however, most resolve spontaneously.
Filamentous or frond-like lesions develop at the teat orifice or on the ventral teat barrel and interfere with milking, either directly by impeding milk flow or indirectly by affecting the teat cup liner closure.

Figure 3.23: Wart in the teat; densely packed (left) and widely dispersed (right)

f. Teat-end hyperkeratosis: Teat-end hyperkeratosis is characterized by hyperplasia of the teat orifice's keratin layer in response to chronic stimuli. Faulty machine milking is the primary reason for it. The teat orifice will show a smooth or roughened raised ring of tissues that later turn into Black spots caused by Fusobacterium necrophorum.

Figure 3.24: Teat end hyperkeratosis

Photo credit: From the web

g. Chaps: Chaps are superficial cuts with abrasions across the teat skins due to damage from a bite from the incisor teeth with a sharp edge. This injury frequently occurs when she-buffaloes are sucked by older calves.

Figure 3.25: Chaps in teat

h. Foot and Mouth disease: Foot-and-Mouth disease is a highly transmissible viral buffalo disease. In the teat, blisters form in the tissue, damaging the cells in the udder. It causes a marked drop in milk production.

i. Intraluminal obstructions: The fibrosis or stenosis in the narrow canal may entirely or partially restrict the teat canal. Visual examination or probe insertion cane is used to identify the obstruction.

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4.1 Common buffalo diseases other than mastitis and their management

Buffalo are less susceptible to pathogens than cattle. Bangladesh has done a few buffalo disease studies. Thus, the country's buffalo disease distribution and prevalence are unknown. Hemorrhagic septicemia (HS), foot and mouth disease (FMD), Black quarter (BQ), brucellosis, and buffalo pox are common infectious diseases that cause economic losses in buffalo.

4.1.1 Bacterial diseases of buffalo

4.1.1.1 Hemorrhagic Septicemia (HS)

HS is an acute, fatal disease of water buffalo in tropical regions caused by specific serotypes of Pasteurella multocida. Infected animals shed the organism during stress, such as high temperature and humidity, concurrent infection, poor nutrition, or work stress. The disease is spread by ingesting contaminated feed or water or by contact with infected oral or nasal secretions from healthy carrier animals or clinically ill animals. The disease is most prevalent during the rainy season.

Clinical sign: The affected animal will die within 8-24 hours in a peracute case. Acute disease is characterized by fever, lethargy, hypersalivation, lacrimation, and nasal discharge. Other symptoms may include abdominal pain, diarrhea, respiratory distress, cyanosis, recumbency, and subcutaneous edema in the pharyngeal area that spreads to the ventral neck and brisket (and occasionally the forelimbs).

Figure 4.1: Clinical signs of HS (a) depression; (b) congested mucous membrane; (c) hypersalivation and (d) edema in submandibular, cervical region towards brisket) (Photo credit: From the web)

Treatment: Antibiotic therapy effectively treats the disease in its early stages. Antibiotics should be chosen based on cultural sensitivity. Symptom-based supportive care, such as NSAIDs, fluid therapy, etc., are provided.

Prevention and Control: Oil adjuvant and Alum precipitated killed HS vaccines (LRI, Bangladesh) have been used to control the disease in the field. Other control measures include animal movement control, culling infected animals, and premise disinfection.

4.1.1.2 Brucellosis

Brucellosis in buffalo is caused by Gram-negative Brucella abortus and B. melitensis. The disease causes abortion (especially in the last trimester), premature births, placenta retention, metritis, infertility, reduced milk production, lameness, epididymitis, and orchitis in buffalo bulls and cows.

The infection spreads through ingestion, conjunctival and intact skin penetration, and udder contamination during milking. Uterine materials, semen, milk, and colostrum are the organism's infection sources.

Disease treatment is not advised. Quarantine the affected animal until all breeding animals have finished gestation. The disease can be controlled by vaccination, limiting animal movement, culling infected buffalo, properly disposing of dead animals, aborted materials, and uterine discharge.

4.1.1.3 Black quarter (BQ)

BQ, caused by Clostridium chauvoei, causes severe lameness and painful, crepitating swelling of the upper leg in buffaloes. Fever, depression, anorexia, dyspnea, rapid heart rate, and prostration at the terminal stage are other symptoms. The disease is transmitted via oral contact with infected wounds and soil, making it a soil-borne disease.

Treatment and prevention: BQ is treated with susceptible antibiotics (such as an increased intravenous penicillin dose), anti-BQ serum, fluid therapy, and supportive therapy. Control measures include vaccination (e.g., LRI BQ vaccine), isolation of diseased animals, and deep burial of carcasses.

4.1.2 Viral diseases of buffalo

4.1.2.1 Foot-and-Mouth Disease (FMD)

Buffalo are less susceptible to FMD than cattle. The disease is caused by aphthovirus. It is spread by inhalation, ingestion, or direct contact. Cattle can pass the virus to buffalo and vice versa. The disease affects all ages, but suckling buffalo calves have a more severe form with high mortality.

Clinical signs include excessive salivation, depression, anorexia, and lameness brought on by painful blisters on the lips, tongue, gums, nostrils, coronary bands, interdigital spaces, and teats. Fever and a significant milk production drop were also observed in affected animals.

Figure 4.2: Nasal mucosal erosions in a buffalo with FMD (Photo credit: From the web)

Buffalo typically recovers faster from the disease. Recovered buffalo can carry the virus for 18-24 months.

Treatment: Lesions are disinfected, emollients are applied, and antibiotics are given to prevent secondary bacterial infections. NSAIDs and honey on mouth lesions are other symptomatic treatments.

Control: Eradication, immunization, or both control the disease. Eradication involves slaughtering all infected and in-contact animals, disinfecting inert materials (clothing, vehicles, machinery) before leaving infected premises, disinfecting infected premises, quarantining the outbreak, epidemiological investigation, etc. Immunization also controls FMD. Killed bivalent and trivalent (A, O, and Asia-1) vaccines are widely available in Bangladesh. Trivalent vaccines are more popular because they contain more antigenically distinct substrains. Local and foreign vaccines are also available.

1.1.2.2 Buffalo pox

The Orthopoxvirus causes Buffalo pox and causes lesions on the teats, udder, and thighs that heal in three to four weeks. Fever, anorexia, dullness, depression, and conjunctivitis are other symptoms. Human infections have been reported, but buffalo infections are not transmissible to cattle.

The only treatment for the disease is symptomatic. By avoiding infected animals and their secretions, the disease can be prevented. Vaccination is also available for buffaloes in other countries but not in Bangladesh. The disease can be prevented by disinfecting udder clothes, milking machines, and milkers' hands.

Figure STYLEREF 1 \s 4. SEQ Figure \* ARABIC \s 1 3: Pox lesions in buffalo udder (Photo credit: From the web)

4.1.2.3 Infectious rhinotracheitis/infectious pustular vulvovaginitis/Red nose

Bovine Herpes Virus 1 (BHV-1) causes Infectious Bovine Rhinotracheitis/Infectious Pustular Vulvovaginitis (IBR/IPV). The virus spreads through aerosols, coitus, and artificial insemination.

The infection causes three symptoms. Fever, decreased milk production, red nose, discharge from the nose and eyes, conjunctivitis, and hypersalivation are symptoms of the respiratory form. Fever, genital mucosa hyperemia, white vulvar discharge, and balanoposthitis are genital symptoms. Fever, salivation, red nose, conjunctivitis, oral mucosa erosions, and respiratory distress are symptoms in new-born calves.

IBR has no treatment. Antibiotics can treat secondary bronchopneumonia. Vaccination against the virus can control the disease. When passive immunity begins to decline, calves should be vaccinated. Bangladesh lacks vaccines for the disease. Farm biosecurity lowers risks.

4.1.3 Parasitic diseases (Endoparasites)

4.1.3.1 Ascariasis

Ascariasis, caused by an adult nematode (Toxocara vitulorum), kills many buffalo calves and causes significant economic loss. The disease affects all ages but is most common in calves under three months. Calves usually get the nematode through the transplacental and transmammary routes. The intestine and other major organs (liver, lungs, etc.) are severely damaged by it.

Clinical signs include intermittent diarrhea, stunted growth, anemia, weakness, moderate dehydration, anorexia, and death, especially in calves between one and three months old.

Figure 4.4: Intestinal obstruction with nematodes in a buffalo calf (Photo credit: From the web)

Treatment and control: Ascariasis is treated with fenbendazole, praziquantel, ivermectin, piperazine, and other supportive therapies. Regular deworming, farm hygiene, biosecurity, and controlling dam egg transmission to calves prevent and control the infestation.

4.1.3.2 Fascioliasis

Fascioliasis, caused by the trematode Fasciola species, is another common buffalo disease that causes weight loss, liver damage, and economic loss. F. hepatica and F. gigantica cause the disease, but the latter is only found in tropical regions like Bangladesh. The parasite's intermediate host is the snail Lymnea.

Clinical sign: Buffalo suffers severe liver damage with chronic Fasciola infection. As a result, animals become anemic, swell in the submandibular region, lose weight, and produce less milk.

Treatment and control: Several anthelmintics, including triclabendazole and albendazole, are effective against the parasite. When combined, triclabendazole and nitroxynil were more effective against immature flukes. Regular deworming, farm hygiene and biosecurity, and snail host movement in the grazing land are necessary to control the infestation.

4.1.3.3 Gigancocotyl (Explanatum) explanatum

explanatum is a trematode parasite commonly found in water buffaloes' bile ducts, gall bladder, and duodenum. Its lifecycle requires two hosts. The definitive host is the water buffalo, while the intermediate host is a snail, Gyrulus convexiculus.
explanatum infection in water buffaloes is common in India and Pakistan. Infection is spread by drinking snail-released cercariae-contaminated water. The parasite can cause liver damage, jaundice, and hemorrhagic duodenitis. Clinical signs of the disease are profuse fetid diarrhea, weight loss, milk production reduction, and death. The animal becomes thirsty and drinks water frequently.

Figure 4.5: G. explanatum (Photo credit: From the web)

Treatment and Control: Oxyclozanide and niclofolan are effective against the worm. Controlling G. explanatum infection in water buffaloes is possible. These include: treating infected animals with an appropriate anthelmintic, treating water with snail-killing chemicals, and raising water buffaloes in areas without the snail host.

4.1.4 Parasitic diseases (Ectoparasites)

4.1.4.1 Tick infestation

Buffalo is considered tick-resistant, so tick infestation is lower than in cattle. Rhipicephalus, Amblyomma, Hyalomma, and Ixodes are the most common buffalo tick species in the Indian subcontinent. They adhere to the host's skin while sucking the blood, which results in anemia, weight loss, pruritus, damage to the skin and hide, and ultimately financial loss.

Topical anthelmintics, manual removal of the parasite from the skin, and destruction of infested bedding and forages are all solutions to the problem.

Rotational grazing, biological control (birds, spiders, ants), acaricides, scraping farm waste (feces, feed waste, etc.) against the walls of unoccupied paddocks, and slow-burning it to kill tick eggs and quarantine of new buffalo are tick infestation control methods.

4.1.4.2 Lice infestation

Another bloodsucking ectoparasite of buffalo is lice. Lice are either sucking (sucking blood and piercing buffalo skin) or chewing (consuming skin cells and debris). Sucking lice cause skin irritation, anemia, anorexia, restlessness, and reduced productivity in buffalo. The most common lice infest buffalo is Haematopinus tuberculatus

Figure 4.7: Lice infestation in buffalo (Photo credit: From the web)

4.1.4.3 Mite infestation (Mange)

Mite infestation in buffaloes causes intense pruritus, anorexia, skin injury (neck, shoulders, axillary area, and udder), and decreased productivity. It lives in the animal's skin. Psoroptes, Chorioptes, and Sarcoptes have been found on buffalo skin. The disease is treated and controlled with agents (permethrin, ivermectin) administered as a subcutaneous injection, spray, pour-on, or dipping. To eliminate mites, treat all animals in a herd, even those without symptoms. Sarcoptes mites infect human skin. Thus, handling a mange animal with caution is advised.

4.1.5 Diseases of mixed infections

4.1.5.1 Calf pneumonia or Bovine Respiratory Disease Complex (BRDC)

In Bangladesh, 90% of buffalo calves die from calf pneumonia. t is a complex, multi-factorial disease caused by Mannheimia haemolytica, P. multocida, Histophilus somni, parainfluenza-3 virus, respiratory syncytial virus, Infectious Bovine Rhinotracheitis Virus, Mycoplasma bovis, and others. When calves are stressed (weather, transportation, stocking density), these organisms cause serious illness.

Clinical signs: Fever, anorexia, rapid shallow respiration, profuse mucopurulent nasal and ocular discharges, and productive cough are common symptoms. A grunting dyspnea may be present.

Treatment and prevention: Antibiotics should be chosen based on cultural sensitivity tests and prescribed with supportive therapy like fluids and NSAID. Preventing pneumonia is the best way to avoid this illness. For example, they provided enough colostrum, lowering stocking density, providing good nutrition, mitigating stress, and practicing good hygiene. The calf pneumonia vaccine is available in several countries but not Bangladesh.

4.1.6 Neonatal diarrheal diseases

It is one of the most crucial buffalo calves’ diseases, causing 10-20% mortality. Poor management infects calves with bacteria, viruses, and endoparasites in their first two months. Bacteria, viruses, and parasites like Eimeria, Strongyloides papillosus, Toxocara vitulorum, and Cryptosporidium cause the disease.

Clinical sign: Affected calves have acute, profuse diarrhea, sometimes with dysentery, cyanotic mucous membranes, depression, weakness, and loss of coordination, which can quickly lead to death within a few days.

Figure 4.8: Buffalo calf with severe diarrhea showing dehydration; sunken eye, eyeball and orbit separation (A), delayed skin fold test (B), eyelids are not closed at all (C), soiling of perineum and tail with yellow feces (D) (Photo credit: From the web)

Treatment and prevention: Treatment with fluids and electrolytes, diet control, and administration of medications based on causal agents. To prevent infection, maintain hygiene, provide enough colostrum, and vaccinate.

4.2 Farm biosecurity

The term "biosecurity" refers to a collection of procedures used to decrease the risk of introducing and spreading disease agents. Adopting attitudes and behaviors to lower risk in all animal care or production operations constitutes implementing Biosecurity.

4.2.1 Farm biosecurity

Farm biosecurity refers to actions used to protect a property against the invasion and spread of pests, diseases, and infestations. Everyone who enters or uses the farmland, including the owner, is responsible for maintaining farm biosecurity.

4.2.2 Importance

Biosecurity is essential in many ways. It improves many aspects at a farm or individual, or regional level, such as,

Improving profit via higher productivity
Obtaining farm stability
Improving the quality and safety of the products
Improving animal welfare
Early detection and immediate reporting of exotic diseases
Reducing antimicrobial use in animals
Reducing zoonoses through preventing and controlling those diseases

4.2.3 Principles of farm biosecurity

To design a biosecurity program, Dewulf & Immerseel (2019) suggested the following principles that are of value in all different environments –

Separation of high and low-risk animals and environments to prevent disease transmission (via quarantine, change of clothing and footwear, and so on)
Keep the infection load low enough to let animals' natural immunity handle it.
Prevent high-risk disease transmission routes first, then low-risk ones
Focus on low-risk actions that are repeated frequently on the farm as they may become risk factors. For instance, while there is a low risk of disease transmission by hand in buffalo, handling occurs frequently. Thus, this event could be significant.
Infections are more likely to occur in larger animal populations and more productive herds. More animals can get and stay infected in a larger herd, raising the infection load. Larger herds have more contact with the outside world, which may increase disease transmission (e.g., feed transport).

4.2.4 Components of Biosecurity

The components of Biosecurity can be classified into two groups – external and internal Biosecurity. These groups are discussed briefly below –

External Biosecurity: External Biosecurity includes all measures required to prevent both the entry of infectious pathogens into farms and the dissemination of infectious agents from farms. Some of the measures are,
1. Location of the farm: Airborne pathogens and vectors are important risk factors for a farm. These pathogens can spread from neighboring farms or slaughterhouses and destabilize farm immunity.
2. Entrance restriction: The farm should have less access to the people since they act as mechanical vectors of several pathogens.
3. Hygiene of transport vehicles: Transport vehicles can spread pathogens into the farm as they are utilized to transfer live animals or to remove carcasses and manure. So, they should be cleaned and disinfected before entering or leaving the farm.
4. Feed safety: Feed and drinking water facilities (as related equipment) should be managed and checked for their quality and safety on the farm because they can introduce pathogens in the herd efficiently.
5. Newly-purchased animal or animal products: The less introduction of new animal or animal products (e.g., semen) to the herd, the higher control of Biosecurity in the farm as they can possess unintended pathogens against which no farm immunity exists. Maintain a quarantine period for newly purchased animals of at least 3-4 weeks, as it can help prevent the spread of new infections in the herd.
6. Vermin and bird control: Rodents, dogs, cats, and birds are reservoir hosts of certain diseases. So, controlling them from the outside entrance or between the farms' compartments is mandatory for the farm owner.
Internal Biosecurity: Internal Biosecurity includes those measures highly connected with farm management and daily practices on the farm. The main components of this category include,
1. Disease management: A farm should manage adequately to ensure good herd health. It includes correctly handling and treating disease with proper diagnosis, isolation, and registration.
2. Stocking density: High stocking density causes stress, increasing the transmission of pathogens and disease susceptibility. So, minimal or ideal stocking density should be maintained for good herd health.
3. Cleaning and disinfection: Cleaning and disinfection are general prophylactic measures to reduce pathogen loads. Thus, it improves overall herd health.
4. Compartmentalization: Several diseases affect different age groups of animals at different levels. It is crucial to keep these groups apart to avoid the transmission of pathogens between the groups.
5. All in/ All out: The all-in/all-out approach makes cleaning and disinfecting the barn between production batches possible while preventing cross-contamination between consecutive production batches. A crucial step in breaking the infection cycle between subsequent production batches is the strict implementation of the concept. This method is best for those large commercial farms.

For efficient management and housing purpose, the young stock can be divided into three groups–

Calves below three months old
Calves from 3-6 months old

Calves from 6-12 months old

It was suggested to have a minimum period of 3 weeks between batches.

4.2.5 Guidelines for cleaning and disinfection (C&D)

Cleaning is the physical removal of organic materials to maximize the exposure of pathogens and microorganisms to the disinfectant. On the other hand, disinfection is the process that involves the destruction of pathogenic organisms associated with inanimate objects, usually by chemical means like 70% ethyl alcohol, hydrogen peroxide, and potassium permanganate.

A general protocol for cleaning and disinfection, described by Immerseel et al. (2019), is given below from the practical aspects –

Dry cleaning: Equipment like a shovel, brush, and scraper should be used to remove all visible dirt after emptying the stable.
Soaking with water and a cleaning material: The surface should be washed with water and a cleaning product like a foam to loosen any organic materials left after dry cleaning.
High-pressure cleaning: Remove the loosened organic material from the surface by rinsing with high pressure (more than 50 atm but less than 120 atm) using hot or lukewarm water so that all organic debris and cleaning agents are removed. Remove the animals from the stable before cleaning as they can spread aerosols with infections.
Drying: The stable must be dried before disinfection since any remaining water will reduce the disinfectant's effectiveness.
Disinfection: Disinfection is carried out after drying to kill most microorganisms. Several disinfectants (e.g., detergents (soap), alcohol, and oxidizing agents like potassium permanganate) are used in this step. Disinfection can be conducted in two methods – wet and dry disinfection.

Wet disinfection is performed using high pressure or a sprinkler. On the other hand, in dry disinfection, a portable or fixed fogger heats a highly concentrated disinfectant before converting it to fog.

Usually, 20-30 minutes of contact with disinfectant and surface is needed for sufficient disinfection.

Drying: After disinfection, the stable should be dried again to avoid animal contact with disinfectant residue. Stable drying is best with sun drying. The off period between batches should be kept.
Efficacy testing: A crucial phase in the process is verifying the effectiveness of the entire process to ensure that everything has been done correctly. This section is well-discussed in the "How to measure farm biosecurity and hygiene level" section.

4.2.6 How to measure farm biosecurity and hygiene level

Their status should be measured accurately and reproducibly to improve farm biosecurity and hygiene. It is mainly examined by measuring the degree of bacterial contamination on surface and air samples.

Bacteriological monitoring

After cleaning and disinfection, bacteriological monitoring is conducted to observe total aerobic bacterial counts (e.g., indicator organisms like E. coli, Enterococcus spp., Salmonella spp., methicillin-resistant Staphylococcus aureus) or specific pathogens. Indicator organisms are found at a higher level at the sample location than the pathogen and are easy to culture and identify. For example,

coli is an indicator organism for fecal contamination and non-enveloped viruses.
Enterococcus is an indicator organism for fecal contamination of surfaces.

Bacteria can be monitored after cleaning and disinfection by following tools –

Agar contact plates (ACP)

ACP measures and quantifies the bacterial load on the surface (e.g., floors, walls, feeding trough). Sampling for the evaluation is done by pressing the plate against the surface and allowing incubation. Then the bacterial colonies are counted (expressed as colony-forming units (CFU) per plate) and scored between 0 to 5 (Table-1)

Score	CFU per plate
0	0
1	1-40
2	41-120
3	121-400
4	> 400
5	Countless

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Advantages of the process include –

Ease of use
No need for further processing after sampling
Pre-made tools are available

Disadvantages can be listed as follows –

Limited sampling surface (25 cm²)
Applied only smooth, firm surfaces
Colony overgrowth
One ACP per specific organism

Swab and rinse sampling

Swab sampling is a direct surface sampling method best suited to restricted work areas such as extensive work surfaces. On the other hand, rinse sampling is an indirect method and best suited for places that are not easily reachable, like pipes, longer tubes, and large tanks. Sometimes both procedures are performed to complete a comprehensive evaluation.

Advantages of the method include –

Larger sampling area than ACP (625 cm²)
More than one bacterial analysis per swab
Can be sampled from surfaces that are difficult to reach

Disadvantages of the method are –

Need laboratory manipulation
No standardized protocol

Visual inspection

In the past, visual examination was the most popular method for gauging cleaning effectiveness. The end outcome of C&D was the absence of any observable biological material on the surface. But this method is biased as a subjective view of cleanliness varies from person to person.

4.2.7 Scoring of Farm bio-security system

A scoring system uses a risk-based scoring tool to quantify the on-farm biosecurity level. Then experts from the veterinary medicine field are asked to weigh the different biosecurity categories and questions on a scale of 0-100 (lowest to highest). The end scores will inform farmers which farm components should be improved and allow comparison between other farms.

An example of a farm biosecurity report is provided below –

Sl. No.	Description	Score (%)
*External Biosecurity*
1	Purchase of new buffalo	80
2	Feed and water supply	85
3	Removal of manure and dead animals	83
4	Entrance of visitors and personnel	90
5	Location of the farm	75
6	Infrastructure	87
7	Control of biological vectors	86
8	Transport vehicle hygiene	70
*Average of scores of external Biosecurity =*		82
*Internal Biosecurity*
1	Disease management	84
2	Cleaning and disinfection	89
3	Compartmentalization	73
4	Distance between compartments	94
5	Stocking density	75
*Average of scores of internal Biosecurity =*		83
Total average =		82.5

4.3 Vaccination and deworming

4.3.1 Vaccination

Regular vaccination against common diseases of buffaloes is one vital way to prevent the spread of some infectious diseases in the herd. Common diseases infecting buffaloes are foot and mouth disease (FMD), hemorrhagic septicemia (HS), black quarter (BQ), and anthrax, which may be prevented using vaccines.

Farmers can follow the below-mentioned vaccination schedule for disease prevention-

Sl no	Vaccine name	Source of vaccine	1^st dose	Bootstring	Dose & Route
1	Anthrax	Government	Six months or more	Yearly	1ml, SC route
2	FMD	Government/ Private	Four months	Four months interval	6ml SC route
3	HS	Government	Six months or more	Yearly	2ml, SC route
4	BQ	Government	Six months	Six months intervals up to 3 years of age	5 ml SC route

Points to be considered during vaccination -

Always vaccinate healthy animals.
Vaccinate the animals in the early morning or evening when the temperature is low.
Use a sterilized disposable syringe for vaccination.
Strictly maintain cold chain during storage and vaccine transport.
Use the vaccine within 2 hours after opening.
Consult with a veterinarian when vaccinating pregnant animals.

4.3.2 Deworming

Regular deworming is vital to maintaining a healthy buffalo herd. It should be started with piperazine citrate at three weeks of age to control neonatal ascariasis. Deworming should be done every month for the first six months of age and, after that, once every three months. Routine deworming of buffaloes every 3-4 months interval is suggested for buffaloes in the free-range, semi-intensive, and semi-bathan systems. In more controlled intensive farms, fecal examination is recommended before deworming therapy.

Points should be considered for deworming –

Deworming should be done every three months intervals.
It can be administered orally (Bolus) or subcutaneously (injection).
Avoid deworming diseased animals.
Multivitamins and liver tonic can be given after deworming.

Piperazine citrate, albendazole, levamisole, cypermethrin, and thiabendazole are commonly used anthelmintics for treating and preventing parasitic infestation in buffalo.

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