Introduction: The aim of the study was to evaluate the occurrence of enterococci in inflammatory secretions from mastitic bovine udders and to assess their antimicrobial resistance. Material and Methods: A total of 2,000 mastitic milk samples from cows were tested in 2014–2017. The isolation of enterococci was performed by precultivation in buffered peptone water, selective multiplication in a broth with sodium azide and cristal violet, and cultivation on Slanetz and Bartley agar. The identification of enterococci was carried out using Api rapid ID 32 strep kits. The antimicrobial susceptibility was evaluated using the MIC technique. Results: Enterococci were isolated from 426 samples (21.3%). Enterococcus faecalis was the predominant species (360 strains), followed by E. faecium (35 isolates), and small numbers of others. The highest level of resistance was observed to lincomycin, tetracycline, quinupristin/dalfopristin (Synercid), erythromycin, kanamycin, streptomycin, chloramphenicol, and tylosin. Single strains were resistant to vancomycin and ciprofloxacin. All isolates were sensitive to daptomycin. E. faecalis presented a higher level of resistance in comparison to E. faecium, except to nitrofurantoin. Conclusion: The results showed frequent occurrence of enterococci in mastitic cow’s milk and confirmed the high rate of their antimicrobial resistance.

Objective: The objective of this study was to investigate the intensity of phage infection caused by Phage SAvB14, which was isolated from dairy farms, depending on the initial number of Staphylococcus aureus cells in the medium. Material and methods: To evaluate the impact of the viable bacteria S. aureus var. bovis on the intensity of phage infection caused by Phage SAvB14, 1 mg of phagolysate (phage titer 105 CFU/ ml) was introduced in 9 ml of nutrient broth with an appropriate amount of daily culture of S. aureus var. bovis under study. The number of viable staphylococci was determined by total viable count/ml. Results: In this experiment, we found that the intensity of phages lytic activity was dependent on the number of sensitive bacterial cells in the volume of the culture medium. Effective phage ther¬apy requires a high concentration of phages in the medium (inflammation foci) for rapid contact of the virus with bacteria. Conclusion: When developing a phage drug to treat subclinical mastitis, it is necessary to increase the phage titer in the drug or its dosage compared to the clinical form, as there is a lower proba¬bility of phage contact with a susceptible microbial cell. Besides, at a high concentration of bacte¬ria, there is a gradual decrease in nutrients in the medium, resulting in phages going back to the condition of lysogeny. [J Adv Vet Anim Res 2020; 7(3.000): 509-513]

Objective: The present study was an attempt to delineate the roles played by the neutrophil surface adhesion molecules and toll like receptors (TLRs) in crossbred cows suffering from Staphylococcus aureus subclinical and clinical mastitis.Materials and methods: Thirty six Karan Fries (KF) cows were categorized into three groups namely healthy (n=12), subclinical mastitis (SCM; n=12) and clinical mastitis (CM; n=12) after screening 146 cows. The grouping was done based on evaluation of collected milk samples by routine procedures like Californian Mastitis Test (CMT) scoring, microscopic counting of milk cells (SCC), bacterial culture of milk samples and observing gross changes in milk. Culture of milk and blood was done for the identification of Staphylococcus aureus subclinical and clinical mastitis. Results: Healthy cows expressed significantly (P<0.05) higher L-selectin (CD62L) in both milk and blood neutrophils as compared to the animals suffering from SCM and CM; however, no significant difference was noticed between milk and blood neutrophils. Significant (P<0.05) increase in the expression of beta integrin (CD11b) was observed in the CM group of cows as compared to SCM and healthy cows. Similar trend in the expressions of TLR2 and TLR4 in both blood and milk neutrophils was observed in the CM cows as compared to the healthy and SCM cows. Milk neutrophils revealed a higher expression of TLR as compared to blood neutrophils.Conclusion: Host elicits stage specific expression of surface adhesion molecules and TLR2 and TLR4 as dynamic host innate immune response against Staphylococcal mastitis. [J Adv Vet Anim Res 2016; 3(2.000): 99-105]

Mastitis is associated with release of free radicals, increased total oxidant capacity and decreased total antioxidants capacity in milk. Antioxidant vitamins and minerals protect the body from free radicals either by directly scavenging free radicals or by inhibiting the activity of oxidizing enzymes. The supplementation of mastitic dairy cows with antioxidant vitamins as vitamin A (VA) and β-carotene (BC), vitamin C (VC), vitamin E (VE), and antioxidant minerals as selenium (Se), Zinc (Zn) and copper (Cu) is very important to help the animal recover early. The aim of this review was to discuss the oxidative stress in dairy cows’ mastitis, and the roles of VA and BC, VC, VE, Se, Zn, and Cu in mastitis of dairy cows. Before deciding to supplement dairy cow rations with the levels of vitamins and minerals, dairy farmers should have their animal feeds tested and their rations evaluated by a competent dairy cow nutritionist and a trustworthy laboratory to be sure what levels of supplementation may be warranted. While inadequate intake and absorption of certain nutrients may result in a weakened immune system and perhaps more mastitis during the lactation period, unjustified supplementation can be expensive and lead to other animal health problems.

Objective: This work aimed to determine the resistance and/or the susceptibility to antibiotics of staphylococci isolated from cattle with mastitis in the North of Algeria. Materials and Methods: The disk diffusion method was carried out to reveal the antibiotic resis¬tance in accordance to the National Committee for Clinical Laboratory Standards guidelines in the Mueller-Hinton agar. Results: Coagulase-negative Staphylococci (CNS) isolates showed more resistance to Cefoxitin, Amoxicillin + Clavulanic Acid, Vancomycin, Trimethoprime Sulfamethoxazole, Clindamycine, Neomycin, and Erythromycin than Coagulase-positive Staphylococci (CPS). CPS were more resis¬tant to Penicillin and Tetracycline as compared to CNS strains; however, all these strains presented sensitivity to Gentamicin and neomycin. Conclusion: The Staphylococci showed high resistance to the beta-lactam antibiotics. As far as the authors know, these molecules are used with or without control in different protocols to prevent and cure the mastitis in Algeria. [J Adv Vet Anim Res 2019; 6(2.000): 231-235]

Objective: The objective of this study was to develop a low-cost kit for the detection of subclinical mastitis (SCM) and to check its validity, reproducibility, and efficacy at the field level. Materials and Methods: A total of 550 quarter milk samples from crossbred dairy cows were collected, of which 400 milk samples were used to validate the newly developed BLRI mastitis test (BMT) kit to justify its efficacy as an individual test kit in detecting SCM based on somatic cell count (SCC) by direct microscopic count (DMC). The efficacy of the newly developed BMT was compared with the California Mastitis Test (CMT) kit. Another 150 milk samples were subjected to SCC determined by DMC and DCC (De Laval cell counter®) categorized by CMT and BMT scores. Results: A SCM test kit, namely, BMT kit was successfully developed in this study. The percentage accuracy of CMT and BMT were 76.75% and 75.75%; sensitivity 69.36% and 67.56%; specificity 85.95% and 85.85%; positive predictive value 86.03% and 85.71%; negative predictive value 69.23% and 68%, respectively. A p value of 0.001 was found for both CMT and BMT. However, CMT and BMT had no significant difference in sensitivity (p = 0.778). Average SCCs (cells/ml) determined by DCC and DMC, respectively, were mostly corresponded to the SCC ranges of both CMT and BMT scores. Conclusion: The newly developed BMT kit is an independent, cheap, farmer-friendly, first country made, and reliable SCM diagnostic test kit that can be used at field condition. [J Adv Vet Anim Res 2019; 6(3.000): 425-430]

Objective: The objective of this study was to identify the multi-drug resistance (MDR) Klebsiella sp. from mastitis milk samples. Materials and Methods: In the current research, 48 clinical mastitis milk samples were collected from Rangpur division, Bangladesh. Confirmation of bovine mastitis (BM) was done by the California Mastitis Test (CMT). All the CMT positive isolates were subjected for the identification of Klebsiella sp. using through a series of cultural and biochemical tests. MDR Klebsiella sp. isolates were determined using the disk diffusion method, and minimum inhibitory zones were measured by following Clinical and Laboratory Standards Institute. MDR patterns of the isolates were also subjected to study by using housefly (Musca domestica). Results: Among the isolates, 62.5% (n = 30/48) revealed the presence of Klebsiella sp. Eight antimicrobial agents including Amoxicillin, Novobiocin, Erythromycin, Vancomycin, Cephradine, Tetracycline, Bacitracin, Methicillin, and housefly (M. domestica) showed complete resistance to Klebsiella sp. On the other hand, Chloramphenicol, Gentamicin, Ciprofloxacin, Azithromycin, Norfloxacin, Levofloxacin, and Nalidixic acid showed sensitivity. Conclusion: This study helps to treat BM with effective antibiotics and helps in an epidemiological study in Rangpur division as well as helps to create public health awareness. [J Adv Vet Anim Res 2019; 6(3.000): 362-365]

Mastitis is an infectious disease condition resulting in inflammatory reaction which occurs when a large number of leukocytes migrate into the mammary gland. Mastitis causes significant financial losses for the global dairy industry. The present study aimed at development of transdermal film comprising of Azadirachta indica extract for treatment of mastitis, which is economical, safe and will not emerge multidrug resistance amongst pathogens. Azadirachta indica extract was prepared by sonication method. The extract was evaluated for minimum inhibitory concentration against gram-positive and gram-negative organisms. Transdermal films with Azadirachta indica extract were prepared with ethylene vinyl acetate and polyvinyl acetate then subjected for the physical characterization, drug content, in vitro dissolution, tensile strength and antibacterial activity. The minimum inhibitory concentration of extract displayed 5mg/ml. Based on quantification of azadirachtin by UV spectrophotometer 100 mg of extract was incorporated in each transdermal film. Transdermal films of Azadirachta indica extract possessed desirable physical properties, and exhibited the drug release of 91.54% in 8 hours. Transdermal film of Azadirachta indica showed sensitivity with a zone of inhibition of 19mm, 16mm, 19mm and 22mm against Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa respectively. Data obtained revealed that transdermal films of Azadirachta indica extract exhibited good drug release profile and desirable physical properties, with good antibacterial activity. The present work reveals that transdermal film of Azadirachta indica extract can be considered for the treatment of mastitis.

Despite of the great efforts to develop effective control programs for mastitis, it is still one of the most economically important diseases in dairy cattle herds. Pseudomonas aeruginosa is a member of coliform Gram-negative bacteria causing treatment-resistant clinical or sub-clinical mastitis in dairy cows. This study aimed to evaluate the influence of affection with clinical P. aeruginosa mastitis on some oxidative stress biomarkers, inflammatory cytokines and proteins, in addition to some complement factors in Holstein dairy cows. Affection with P. aeruginosa mastitis evoked a state of oxidative stress which accompanied with depletion of cellular enzymatic and non-enzymatic anti-oxidants and elevation of lipid peroxide and advanced oxidation protein product (AOPP) level. Additionally, this affection stimulated the release of some inflammatory cytokines and proteins, enhanced activity of caspase-1. In contrary, the level of complement factor 2 (C2), complement fragments C3b and complement fragment C5a has been decreased upon affection with mastitis. In conclusion, marked oxidative stress state and enhanced release of inflammatory cytokines and proteins with complement system defective activation may share in pathogenesis and virulence of P. aeruginosa-induced clinical mastitis in dairy cattle.

Objective-To investigate effects of intramammary administration of prednisolone on the immune response of mammary glands in cows. Animals- 5 lactating Red Holsteins. Procedures- Cows received a different intramammary infusion in each mammary gland (10 mg of prednisolone, 100 μg of lipopolysaccharide [LPS], 100 μg of LPS and 10 mg of prednisolone, or saline [0.9% NaCl] solution). Milk samples were collected before (time 0) and 3, 6, 9, 12, 24, and 36 hours after treatment. Somatic cell count (SCC), lactate dehydrogenase (LDH) activity, and concentrations of serum albumin (SA) and tumor necrosis factor (TNF)-α in milk and mRNA expression of TNF-α, interleukin (IL)-8, and IL-1β in milk somatic cells were analyzed. Results-Saline solution or prednisolone did not change SCC, LDH activity, and SA and TNF-α concentrations in milk and mRNA expression of TNF-α, IL-1β, and IL-8 in milk somatic cells. The SCC and TNF-α concentration in milk increased similarly in glands infused with LPS, independent of prednisolone administration. However, the increase of LDH activity and SA concentration in milk after LPS infusion was diminished by prednisolone administration. The mRNA expression of TNF-α, IL-8, and IL-1β in milk somatic cells increased after LPS infusion and was unaffected by prednisolone. Conclusions and Clinical Relevance- Intramammary administration of prednisolone did not induce an immune response and did not change mRNA expression of TNF-α, IL-8, and L-1β during the response to intramammary administration of LPS. However, prednisolone reduced disruption of the blood-milk barrier. This could influence the severity and cure rate of mastitis.