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Mammalian milk contains a variety of complex bioactive and nutritional components and microorganisms. These microorganisms have diverse compositions and functional roles that impact host health and disease pathophysiology, especially mastitis. The advent and use of high throughput omics technologies, including metagenomics, metatranscriptomics, metaproteomics, metametabolomics, as well as culturomics in milk microbiome studies suggest strong relationships between host phenotype and milk microbiome signatures in mastitis. While single omics studies have undoubtedly contributed to our current understanding of milk microbiome and mastitis, they often provide limited information, targeting only a single biological viewpoint which is insufficient to provide system-wide information necessary for elucidating the biological footprints and molecular mechanisms driving mastitis and milk microbiome dysbiosis. Therefore, integrating a multi-omics approach in milk microbiome research could generate new knowledge, improve the current understanding of the functional and structural signatures of the milk ecosystem, and provide insights for sustainable mastitis control and microbiome management., Competing Interests: Declarations. Competing interests: The authors have no relevant financial or non-financial interests to disclose. Ethical approval: No ethical approval is required. Consent to participate: Not applicable. Consent to publication: Not applicable., (© 2025. The Author(s).)

Camel mastitis especially caused by Staphylococcus aureus (S. aureus), is a major risk to animal health and milk production. The current investigation evaluated the antibiotic susceptibility and virulence factors of S. aureus isolates from subclinical mastitis in camels. A total of 384 milk samples were collected and submitted to isolate S. aureus. The S. aureus isolates exhibiting resistance to Penicillin and Cefoxitin disc on Kirby-Bauer disc diffusion method were considered as β-lactam resistant S. aureus (BRSA) and methicillin-resistant S. aureus (MRSA) which were further confirmed by PCR targeting blaZ and mecA genes, respectively. The results showed that S. aureus was found in 57.06% of subclinical (SCM) positive camel milk samples. A high molecular prevalence of BRSA and MRSA were found to be 48.51% and 46.53% respectively depicting that treating these infections is challenging due to their high resistance levels. The phylogenetic analysis revealed a significant resemblance of the study isolates with each other and with already reported sequences from different countries which shows the potential for the spread of pathogen. Virulence profiling of antibiotic resistance strains showed the presence of virulence markers (nuc and coag genes), intercellular adhesion genes (icaA, icaD), Panton-Valentine leukocidin (pvl) gene, and enterotoxin-producing genes including sea, seb, sec, and sed. In-vitro antibiotic susceptibility testing revealed that the most resistant antibiotic group was penicillin followed by aminoglycosides and cephalosporins. Drug repurposing analysis of different non-antibiotics for combination therapies with resistant antibiotics was done to combat the S. aureus isolates harboring the mecA and blaZ genes. The results revealed the synergistic effect of amoxicillin, sulfamethoxazole, gentamicin, and doxycycline with ketoprofen, amikacin with flunixin meglumine, and gentamicin with N-acetylcysteine (NAC) against study isolates. The current investigation provides the status of antibiotic-resistant strains and virulence factors of S. aureus in the udder of dromedary camels. The combinational therapy of resistant antibiotics with non-antibiotics provides a potential therapeutic option for the treatment of resistant strains., Competing Interests: Declarations. Ethical approval: Not required. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The present study investigates the prevalence and etiology of subclinical mastitis in Țurcana sheep flocks located in south-western Romania. Milchtest and California Mastitis Test (CMT), were used for the detection of subclinical mastitis. A number of 360 milk samples across three lactation phases were analyzed. Subclinical mastitis was identified in 146 (40.6%) of sheep milk samples. Only milk samples (n = 146) collected from sheep diagnosed with subclinical mastitis were considered for microbiological analyses. Thus, Staphylococcus aureus (129/146; 88.4%) was identified as the predominant pathogen followed by Streptococcus spp. (14/146; 9.6%) Enterococcus spp. (7/146; 4.8%), Pseudomonas aeruginosa (6/146; 4.1%) and Klebsiella pneumoniae (5/146; 3.4%). Antimicrobial susceptibility of isolated strains of Staphylococcus aureus, Streptococcus spp. and Enterococcus spp. was also tested. The results showed varying resistance patterns depending on the farm and microorganism. In heard A the highest resistance of Staphylococcus aureus was identified to polymyxin B (79.7%) followed by novobiocin (73.4%) and tetracycline (45.3%); Streptococcus spp. to streptomycin (100%), novobiocin (100%), tetracycline (100%) and Enterococcus spp. to tylosin (100%), streptomycin (75.0%) and tetracycline (75.0%). In herd B, Staphylococcus aureus showed high resistance rates to polymyxin B (86.3%), novobiocin (74.5%) and cloxacillin (39.2%); Streptococcus spp. strains to streptomycin (100%), novobiocin (100%), tetracycline (100%) and Enterococcus spp. to gentamicin (100%), streptomycin (66.7%) and erythromycin (66.7%). The obtained preliminary results indicate a potential risk that future treatments of sheep mastitis may not be effective, and demonstrated that raw unpasteurized milk can harbor antimicrobial resistance pathogens posing a threat to public health. However, further investigations involving molecular research on resistance genes are required to draw comprehensive conclusions., Competing Interests: Declarations. Ethical approval: Even though the research entitled “Etiology and Antimicrobial Resistance of Subclinical Mastitis Pathogens Staphylococcus aureus, Streptococcus spp. and Enterococcus spp. in Sheep Milk” included live animals (ovines), however, they were not directly affected or used because the samples processed and analyzed in the study were represented by milk samples collected from sheep during milking stages. The statement was included in the Bioethics Commission Approval No. 5639. The experiments assure the proper accommodation and animal handling in accordance with the Directive 2010/63/EU. Also the experimental protocol includes methods for preventing and reducing animal distress in accordance to the recommendation of European Union and Romanian Legislation. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Mastitis is one of the most serious diseases that threatens the health of dairy animals. The somatic cell count (SCC) in milk is widely used to monitor mastitis. This study aimed to reveal the diversity of microorganisms in buffalo milk with high somatic cell count (SCC ≥ 3 × 10 5 cells/mL, n = 30) and low somatic cell count (SCC ≤ 5 × 10 4 cells/mL, n = 10), and identify the dominant bacteria that cause mastitis in a local buffalo farm. We also investigated the potential method to treat bacterial mastitis. The V3-V4 region of 16 S rDNA was sequenced. Results showed that, compared to the milk with low SCC, the high SCC samples showed lower microbial diversity, but a high abundance of bacteria and operational taxonomic units (OTUs). By in vitro isolation and culture, Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae were found to be the leading pathogens, which is consistent with the 16 S rDNA sequencing data. We further isolated 3 of the main pathogens and established a pathogen detection method based on ELISA. In addition, the antibacterial effects of 10 antimicrobials and 15 Chinese herbal extracts were also investigated. Results showed that the microbial has developed tolerance to several of the antimicrobials. While the water extracts of Chinese herbal medicine such as Galla Chinensis, Coptis chinensis Franch, Terminalia chebula Retz, and Sanguisorba officinalis L can effectively inhibit the growth of main pathogens. This study provides novel insight into the microbial diversity in buffalo milk and a reference for the prevention, diagnosis, and treatment of mastitis., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

In herds of dairy goats, mastitis represents a major health and economic problem due to the multiresistance of some microorganisms. In this context, the study aimed to determine the potential of antimicrobial action and antibiofilm of the crude ethanolic extract (CEE) of Hymenaea martiana (jatobá) leaves, as well its fractions, on Staphylococcus sp isolated from bacterial cultures of goat milk. In vitro assays were performed to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC), as well as tests of the effect of CEE on biofilm formation and quantification and the consolidated biofilm. The experimental infection was performed in two groups, each consisting of five goat. Experimental Group 1 (G1) consisted of five females treated with an intramammary ointment based on the CEE, at a concentration of 5%. Experimental Group 2 (G2) consisted of five females treated with a commercial intramammary ointment based on gentamicin, once a day, for six consecutive days. The diagnosis of mastitis was performed using a bacterial culture. The dichloromethane fraction of CEE was the one with the lowest concentrations of MBC, ranging from 195.3 to 781 µg / ml. Concerning to the biofilm, interference of the tested extract was observed for two isolates. In the present study, the ointment prepared from H. martiana extract (jatobá) was able to reduce bacterial infection in mammary glands experimentally infected with S. aureus. Antibacterial activity may be related to the classes of secondary metabolites found., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Lactococcus garvieae (L. garvieae) is a pathogenic bacterium that is Gram-positive and catalase-negative (GPCN), and it is capable of growing in a wide range of environmental conditions. This bacterium is associated with significant mortality and losses in fisheries, and there are concerns regarding its potential as a zoonotic pathogen, given its presence in cattle and dairy products. While we have identified and characterized virulent strains of L. garvieae through phenotyping and molecular typing studies, their impact on mammary tissue remains unknown. This study aims to investigate the pathogenicity of strong and weak virulent strains of L. garvieae using in vivo mouse models. We aim to establish MAC-T cell model to examine potential injury caused by the strong virulent strain LG41 through the TLR2/NLRP3/NF-kB pathway. Furthermore, we assess the involvement of NLRP3 inflammasome-mediated pyroptosis in dairy mastitis by silencing NLRP3. The outcomes of this study will yield crucial theoretical insights into the potential mechanisms involved in mastitis in cows caused by the L. garvieae-induced inflammatory response in MAC-T cells., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The present study aimed to provide a detailed characterization of coagulase-negative staphylococci (CoNS) isolated from cows and buffaloes with mastitis. The study included seventy-five CoNS isolates (60 came from cattle and 15 from buffaloes) originating from 68 individual quarters of 67 dairy cows (53 cattle and 14 buffaloes). The animals belonged to five different small holding dairy herds (n = 140 cows) that show clinical or subclinical mastitis. CoNS isolates were phenotypically characterized using MALDI-TOF-MS and were further genotypically characterized by microarray-based assays. Furthermore, the antimicrobial susceptibility of CoNS strains which carried the mecA gene was examined by broth microdilution. The occurrence of CoNS in the respective five herds was 10.5%, 14.7%, 14.8%, 12.8%, and 9.9%, with an average of 12.4%. Six different CoNS species were identified: S. sciuri (n = 37; 30 from cattle and 7 from buffaloes), S. chromogenes (n = 14; 8 from cattle and 6 from buffaloes), S. haemolyticus (n = 10; nine from cattle and one buffalo), S. xylosus (n = 10; nine from cattle and one buffalo), S. hyicus (n = 2), S. warneri (n = 1), and unidentified CoNS (n = 1). Twenty percent (20%) of CoNS isolates (17.3% of cattle origin) carried at least one antimicrobial resistance gene, while 4% of the isolate including two isolates of S. haemolyticus and one S. warneri of cattle origin carried the mecA gene and were phenotypically identified as methicillin-resistant strains. The genes detected were blaZ (16%), followed by tet(K) (8%), aacA-aphD (4%), aphA3 (2.6%), msr(A) (2.6%), [far1 (2.6%), and fusC (2.6%)], sat (2.6%), and cat (1.3%) conferring resistance to penicillin, tetracycline, gentamicin, neomycin/kanamycin, erythromycin, fusidic acid, streptothricin, and chloramphenicol, respectively. The majority of investigated CoNS strains displayed considerably low prevalence of resistance genes, while resistance to more than three antibiotics was found in S. haemolyticus and S. warneri. Implementing effective preventive measures is, therefore, important for limiting the transmission of CoNS, rather than using antibiotics to control mastitis in bovines.

The present study determined trends in the isolation of bacterial pathogens in milk samples submitted from clinical mastitis buffaloes at the University mastitis laboratory during 2007 to 2016. Culture results (n = 3945) were classified into six categories (Staphylococcus spp., 28.7%; Streptococcus spp., 1%; Corynebacterium spp., 1.1%; gram-negative organisms, 3.2%; others including mixed growths, 3.3%; and no growth, 62.7%). Overall, culture results were significantly associated with year (χ 2  = 112.28; df = 10; P < 0.001) and season (χ 2  = 136.03; df = 20; P < 0.001). From 2007-2009 to 2013-2016, the isolation of Staphylococcus spp. increased from 28.2 to 48.4%, while isolation of Streptococcus spp. (50 to 20%) and Corynebacterium spp. (65.1 to 16.3%) decreased. The proportion of samples characterized as no growth increased from 26.4% (2007-2009) to 46.3% (2013-2016). Further, the proportion of clinical mastitis cases increased over the study years (goodness-of-fit χ 2  = 59; df = 2; P < 0.001). With respect to season, the proportion of Staphylococcus spp. (30.6%; 346/1132), Corynebacterium spp. (48.8%; 21/43), and gram-negative bacteria (55.9%; 71/127) were more associated with the monsoon, and that of Streptococcus spp. (47.5%; 19/40) with the winter. Seasonal effects were also observed on the occurrence of clinical mastitis cases; comparatively, more clinical mastitis cases (35.6%; 523/1473) were observed during the monsoon season (goodness-of-fit χ 2  = 276.7; df = 4; P < 0.001) as compared with other seasons. The present study concludes that Staphylococcus spp. are the major cause of intramammary infections in buffaloes with clinical mastitis, demanding more studies regarding its epidemiology and pathogenicity. Also, more efforts are needed in the prevention and control of clinical mastitis in buffaloes, especially, during the monsoon season.

The present study aims to evaluate the amount of immunoglobulins A, G, and M in she-camel blood serum in relation with the presence of pathogens in the udder, and to compare the antioxidative capacity and the concentration of zinc, iron, total proteins, and calcium. Milk and blood samples from she-camels from south Jordan were taken; according to milk bacteriological examination, the animals were divided into two groups: (Gm) which contained samples of milk contaminated with bacteria and (Gh) which contained uncontaminated milk samples. Milk and blood were sampled from 30 females and examined for the concentration of immunoglobulins A, G, and M and for the presence of pathogens in milk. Total antioxidant capacity, zinc, iron, total proteins, and calcium concentrations in blood were determined. Milk samples were checked for the presence of pathogens. She-camels for the study were of similar age and productivity in the middle stage of lactation. It was determined that the presence of pathogenic bacteria infecting the udder quarters had considerably influenced the values of immunoglobulins G, A, and M, total antioxidant capacity, and zinc and total protein concentration (p < 0.05) in blood serum; no significant difference in iron and calcium concentration was determined. Subclinical mastitis has a crucial role in increasing the concentration of immunoglobulins in serum; some parameters measured in blood (zinc, total antioxidant capacity, total proteins) could be indicative for the presence of inflammation in she-camels.

The objective of this study was to evaluate different screening limits for the California mastitis test (CMT) and the somatic cell count (SCC) in previous diagnoses of subclinical mastitis in Santa Inês and Morada Nova ewes, which were reared under the same management conditions. Additionally, cutoff points were defined for SCC in accordance with the sensitivity and specificity of the test. A total of 907 mammary halves were subjected to CMT and SCC. The disease was confirmed by means of microbiological identification. Coagulase-negative staphylococci (CNS) were the microorganisms with highest occurrence. The CMT score of 1+ provided adequate sensitivity and specificity at all periods of lactation investigated. This score showed good agreement with SCC, >400,000 cells mL(-1). Higher cell counts favored higher diagnostic specificity. They can be used when producers have financial difficulties relating to treatment or culling of sheep with subclinical mastitis. However, producers should be warned about the risk of false-negative results in the flock.

Mastitis caused by microbial infections in dairy goats reduces milk yield, modifies milk composition, and potentially contributes to morbidity in herds and consumers of dairy products. Microorganisms associated with mastitis in dairy goats are commonly controlled with antibiotics, but it is known that continued use of these chemical agents promotes antibiotic resistance among bacterial populations. Recently, it has been shown that bacteriocins of Bacillus thuringiensis inhibit growth of food-borne pathogens and also bacteria associated with bovine mastitis. However, there is no report on their ability to inhibit microorganisms linked to mastitis in dairy goats. In this study, using 16S rDNA and ITS regions of rDNA, we identified nine bacterial isolates and an encapsulated yeast associated with mastitis in dairy goats. Enterococcus durans, Brevibacillus sp., and Staphylococcus epidermidis 2 were resistant to, respectively, 75, ~67, ~42, and ~42 % of the antibiotics screened. In addition, 60 % of the bacterial isolates were resistant to penicillin, ampicillin, vancomycin, and dicloxacillin. Importantly, 60 % of the isolates were inhibited by the bacteriocins, but S. epidermidis 1, Enterobacter sp., Escherichia vulneris, and Cryptococcus neoformans were not susceptible to these antimicrobial peptides. Using Brevibacillus sp. and Staphylococcus chromogenes as indicator bacteria, we show that peptides of ~10 kDa that correspond to the molecular mass of bacteriocins used in this study are responsible for the inhibitory activity. Our results demonstrate that multiple antibiotic-resistant bacteria associated with subclinical mastitis in dairy goats from Guanajuato, Mexico, are susceptible to bacteriocins produced by B. thuringiensis.

Twenty-one dairy ewe flocks selected by stratified random sampling were subjected to study the prevalence and etiology of subclinical intramammary infections and to assess the influence of parity on the prevalence of intramammary infections. Also, spontaneous cure rates were determined over study period. A total of 1192 milk samples were collected at 2 weeks after lambing until tenth-week postpartum. All flocks had hand milking; those which were classified by bacterial culture and California Mastitis Test (CMT) as positive were deemed to have glands with subclinical mastitis (SCM). Of 1192 halves examined, 791 samples were collected during spring and 401 samples were collected during summer. Prevalence rate of SCM in spring was 14.7 %; and spontaneous cure that occurred in this season was 88.8 %; coagulase-negative staphylococci (CNS) were the most common isolates (66.6 %). Samples collected in spring showed higher prevalence rate of SCM than summer samples. This rate was 8.9 % in summer. Spontaneous cure rate in this season was 69.4 %, and Staphylococcus aureus (72.2 %) was the most common isolates. SCM was seen at significantly lower rates in left half than in right one (p < 0.05). Multiparous ewes had significantly higher (p < 0.05) SCM prevalence rates than primiparous ewes. The incidence of clinical mastitis (defined as number of clinical cases per 100 ewe-months) was 0.21 and 0.74 in spring and summer, respectively. The isolates from clinical cases in spring were fungi and, from summer, were S. aureus. Also, S. aureus SCM cases were not significantly severe than other SCM cases. In conclusion, multiparous ewes were most at risk, and severity of infection was higher in summer.

Mastitis is the inflammation of the mammary gland. Recent research has shown that Angiopoietin-like protein 2 (ANGPTL2) is a key inflammatory mediator. In the present study, we tested whether there is a correlation between increased ANGPTL2 expression and inflammation in response to Staphylococcus aureus in murine mastitis and the mechanisms involved. Thirty mice were divided into two groups: blank control group, challenged group. The entire infused mammary glands were removed to observe the changes of histopathology, myeloperoxidase (MPO) activity, production of tumour necrosis factor-α (TNF-α) and interleukin (IL)-6, and genes expression of ANGPTL2, TNF-α and IL-6. In challenged group, the structure of mammary glands was damaged and the large areas of cell fragments were observed. The MPO activity, IL-6 and TNF-α concentrations, ANGPTL2, IL-6, and TNF-α mRNA levels were significantly elevated in challenged group compared with blank control group. The present findings indicate ANGPTL2 may mediate the inflammation in murine mastitis through the activation of IL-6 and TNF-α.

The present study was carried to ascertain the association of various risk factors of mastitis in water buffaloes. The milk samples from buffaloes were collected and screened through California Mastitis Test for the presence of mastitis. In the present study, 15.2 % prevalence of subclinical mastitis was recorded both at the government (13.4 %) and private farms (15.5 %). The chi-square analysis showed significantly higher involvement of the right rear and front quarters. The analysis of variance technique showed significant difference in live body weight, milk yield, teat end to floor distance (P < 0.001), udder depth, teat length, and teat diameter in mastitic and healthy buffaloes. The frequency analysis also revealed significant difference between various groups including lactation stage, teat and/or udder pathology, teat shape, and udder shape (P < 0.001). The logistic regression analysis revealed significant positive association of mastitis with milk leakage, live body weight, milk yield, parity, calf suckling, pendulous udder, number of attendants at the farm, dirty hind legs, and udder depth.

A cross-sectional study was carried out from November 2010 up to April 2011 to estimate mastitis prevalence and associated risk factors and to assess its bacterial causes in traditionally managed camels in Borana Zone, Southern Ethiopia. Thus, 348 lactating camels were examined clinically, and subclinical cases were checked with California mastitis test (CMT). The overall prevalence of mastitis was 44.8 % (156/348), comprising clinical (19, 5.4 %) and subclinical (137, 39.4 %) cases. The quarter level prevalence of mastitis was 24.0 % (334/1,392). Of the total 1,392 examined teats, 30 were blind, and hence, from the 1,362 non-blind CMT-examined teats, 22.3 % (304/1,362) were CMT positive. Of the 304 CMT-positive samples, 264 were culture positive (197 Gram-positive, 41 Gram-negative, and 26 mixed isolates), and 40 were culture negative. The prevalence of Staphylococcus aureus was found to be the highest at both the animal (12.8 %, 39/304) and quarter level (2.9 %, 39/1,362). Regression analysis revealed higher likelihood of mastitis occurrence among camels from Dharito (OR = 3.4, 95 % confidence interval (CI) = 1.8, 6.4), Gagna (OR = 3.4, 95 % CI = 1.8, 6.5), and Haro Bake (OR = 2.6, 95 % CI = 1.3, 5.1) than camels from Surupha. Likewise, there was higher chance of mastitis occurrence among camels at the early lactation stage (OR = 2.3, 95 % CI = 1.1, 4.6) and camels with udder/teat lesions (OR = 13.7, 95 % CI = 1.7, 109.4) than among camels at late lactation stage and camels with healthy udder/teats, respectively. In conclusion, this study reveals the current status of camel mastitis in Southern Ethiopia.

A cross-sectional study was conducted in desert environment of Jhang (Pakistan) from November 2008 to October 2009 on she-camels kept under pastoralist conditions to determine the prevalence of mastitis, impact of risk factors, and isolate the dominant mastitis-causing bacteria on total of 150 lactating she-camels by using clinical examination and surf field mastitis test. From the 150 she-camels examined, 69 (46%) were positive for mastitis at animal level, 12 (8%) clinical, and 57 (38%) subclinical. Age, parity number, stage of lactation, breed, production system, hygiene of milking process, and presence of lesion on udder/teat were found significantly associated (p<0.05) with the prevalence of mastitis in she-camels. There was the lowest prevalence (33.33%; 15 of 45) of mastitis in she-camels of 5-7 years of age, while the highest (80%; 12 of 15) in the animals aged between 14 to 16 years. Stage of lactation significantly affected (p<0.05) and was found to be associated with the prevalence of mastitis being the highest (54.55%; 18 of 33) during the initial stage of lactation (0 to 1 month) followed by last 2 months (10-12 months) as 54.17% and mid-stages (1-3 and 3-10 months) of lactation as 28.57% (6 of 21) and 37.50% (9 of 24), respectively. According to breed of camels, it was noted that the prevalence of mastitis affected significantly (p<0.05) being the highest in crossbred (Desi × Mareecha) as 51.39% (37 of 72) followed in order by Mareecha and Desi as 43.14% and 37.04%, respectively. Staphylococcus (42.19%) and Streptococcus (15.63%) genera were the dominant isolates identified. Good hygiene in milking process, milking clinically infected she-camels at last, culling chronic mastitis carriers, treating clinically infected she-camels, and dry period therapy could reduce the prevalence of contagious mastitis in the study area.

Present study is aimed to evaluate the PCR test for detecting the major pathogens of bubaline mastitis (BM) directly from the mastitic milk samples of the buffaloes. Cell lysates of the enriched mastitic milk samples were directly used as template DNA in PCR and the reactivity of genus and/or species specific primers against the selected pathogens of BM was tested in individual simplex PCR assays. Out of the 60 mastitic milk samples tested 30 were positive for E. coli, 21 were positive for S. aureus, 4 were positive for S. agalactiae, 9 were positive for S. dysgalactiae and 7 were positive for S. uberis. Certain samples were positive for more than one pathogen thus indicating the mixed infection of the udder. Although certain mastitic milk samples reacted with Staphylococcus genus specific primers, they didn't react with S. aureus specific primers in PCR, which indicates the implication of other species of Staphylococcus in BM. The processing of mastitic milk samples was also simplified by eliminating the use of expensive reagents. The detection limits of PCR with the cell lysates are sensitive enough for PCR to be used as diagnostic tool for identifying the pathogens of BM.

Antioxidant, antiinflammatory and phagocytic activities were studied in milk polymorphonuclear cells (PMNs) isolated from healthy buffaloes (group I) and during clinical mastitis with the treatment of Enrofloxacin alone (group II) and combined treatment with Enrofloxacin and Vitamin E plus selenium (group III). On days 0,3, 8 and 15 the milk Somatic cell count (SCC) were significantly higher in mastitic milk than in milk obtained from healthy buffaloes. In group II SCC decreased significantly on day 3 and day 8, however in group III reduction in SCC was observed on day 3, day 8 and day 15 (P < 0.05). The antiinflammatory activity was evaluated by determining nitrite plus nitrate (NOx) production in the milk PMNs before treatment and on day 8. NOx activity was significantly higher in mastitic milk than from healthy controls, both before and after treatment (P < 0.05). In group II and group III the activity decreased significantly on day 8 (P < 0.05). The Glutathione peroxidase (GSH-Px) activity was estimated in the milk polymorphonuclear cell (PMNs) supernatant. GSH-Px activity was significantly lower in mastitic buffaloes than in healthy controls, both before and after treatment (P < 0.05). In group II levels did not change in response to treatment, whereas in group III levels had increased significantly on day 8 (P < 0.05). The phagocytic activity (PA) (percentage of neutrophil that had phagocytosed 1-6 bacteria) and phagocytic index (PI) (average number of bacteria/ leukocytes counted in 100 cells) of the milk PMNs was significantly lower in mastitic buffaloes (P < 0.05). In group II the PA and PI did not change in response to treatment, whereas in group III both the parameters had increased significantly on day 8 (P < 0.05). The results of the present experiment indicated enhancement of antioxidative and cellular defense and reduction of somatic cell count in the mastitic animals treated with Enrofloxacin and Vitamin E plus Selenium as compared to the Enrofloxacin treatment alone. Hence Vitamin E plus selenium therapy may be added along with the antibiotics for effective amelioration of intramammary infection in buffaloes.

This report was delineated to study the clinical, bacteriological and therapeutic aspects concerned with acute coliform mastitis in buffaloes. Bacteriological examination of 80 quarter milk samples obtained aseptically from 56 buffaloes with acute mastitis revealed that coliform bacteria was the most common pathogen (45 cases) followed by Staphylococcus aureus (seven cases) then Streptococcus uberis (three cases), and Streptococcus agalactiae (one case). Clinically, hotness, swelling and painful reaction with serous excretion containing clots was recorded in buffaloes with coliform mastitis. The efficacy of ceftiofur was evaluated in the treatment of buffaloes with acute coliform mastitis. Parenteral ceftiofur neither improved clinical signs nor returned milk to pre-infection production level, whereas intramammary ceftiofur and combination of intramammary with parenteral ceftiofur improved the clinical signs in 10/15 and 12/15 buffaloes, respectively. On quarter level, 3/17, 12/17 and 15/21 quarters recovered in groups received parenteral, intramammary and combination therapy, respectively. This study demonstrates that systemic ceftofur is not effective in the treatment of clinical coliform mastitis in buffaloes.

A double-blind, placebo-controlled study was conducted to compare two vaccines using different adjuvants with regard to their ability to stimulate antibody production against the alpha- and beta-toxins and the exopolysaccharide of Staphylococcus aureus. The vaccines contained identical antigens, consisting of inactivated whole bacteria of two strains of S. aureus in addition to alpha- and beta-toxoid. One vaccine contained mineral oil, while the other used a water-soluble acrylic acid polymer resin (Carbopol) as adjuvant. Saline served as the placebo. One hundred and forty ewes were vaccinated twice before lambing, by subcutaneous injection with vaccine or placebo in the region of the supramammary lymph node, and were observed and sampled over a period of 6 months. The vaccine containing mineral oil as adjuvant induced significantly greater immune responses to the alpha- and beta-toxins than did the vaccine containing Carbopol. The latter vaccine induced higher levels of antibodies to exopolysaccharide. The degree of local adverse reactions did not differ between the two groups. The results indicate differences between the oil-adjuvanted and Carbopol-adjuvanted vaccines with regard to their ability to stimulate antibody production against S. aureus protein antigens in sheep.

The right mammary gland of 12 lactating goats was inoculated intracisternally with 1 ml of Mycoplasma capricolum subsp. capripneumoniae (Mcc) containing 10(6) colony-forming units (CFU), while their left mammary halves received 1 ml of sterile PPLO broth only. Two goats served as uninfected controls. The clinical mastitis that developed in the infected mammary halves within 24 h was initially acute but became increasingly chronic by the end of the experiment at 24 days post inoculation (DPI). The disease was characterized by atrophy of the infected mammary halves, leading to marked agalactia and an increase in somatic cell counts, with a preponderance of neutrophils initially and lymphocytes later. The Mycoplasma was re-isolated from infected mammary secretions up to 16 DPI but not from blood. Histopathology revealed that the mastitis was acute and purulent initially, followed by infiltration of lymphonuclear cells and fibroplasia in the lymphomononuclear cells and fibroplasia in the interacinar tissue, and later by massive fibrosis. Immunohistology demonstrated the presence of Mycoplasma-like bodies localized mainly on the surface of acinar/duct epithelial cells. The studies showed that Mcc was highly pathogenic in the caprine mammary gland.

Milk samples (n = 160) from 7 clinically healthy bactrian camels were cultured to detect subclinical udder infection. The samples were assessed by the Californian mastitis test (CMT) and somatic cell count (SCC). Bacteria were recovered from 36 (22.5%) of the milk samples. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were the main organisms found. Infected quarters had significantly higher mean values for the SCC (p < 0.01) and CMT (p < 0.001) than non-infected quarters. All 7 camels were infected with CNS but only 4 with S. aureus. CMT values for S. aureus-infected camels were significantly higher than for those only infected with CNS. The values for SCC and CMT were significantly influenced by the stage of lactation (p < 0.05). No significant difference was found from the effect of the quarters. Both SCC and CMT were of value in predicting the infection status of the udder.

Three cases of deep soft-tissue infections caused by Streptococcus pneumoniae are presented. All patients were previously healthy adults. The first case was a man with a protracted illness in whom pelvic and inguinal abscesses developed at the site of a scar from a traumatic injury several years earlier. The second patient, a woman, had mastitis with systemic symptoms. The third patient was a woman who developed a gluteal abscess after an intramuscular injection of a contraceptive. Cellulitis and deep soft-tissue infections caused by Streptococcus pneumoniae are uncommon, but may occur even in immunocompetent adults.