Mastitis Research Articles

Acute phase protein expressions in secretory and cistern lining epithelium tissues of the dairy cattle mammary gland during chronic mastitis caused by staphylococci

BackgroundMastitis is the most common disease in dairy cattle and the costliest for the dairy farming industry, as it lowers milk yield and quality. Mastitis occurs as a result of interactions between microorganisms and the individual genetic predispositions of each animal. Thus, it is important to fully understand the mechanisms underlying these interactions. Elucidating the immune response mechanisms can determine which genetic background makes an animal highly resistant to mastitis. We analyzed the innate immune responses of dairy cows naturally infected with coagulase-positive staphylococci (CoPS; N = 8) or coagulase-negative staphylococci (CoNS; N = 7), causing persistent mastitis (after several failed treatments) vs. infection-free (i.e., healthy [H]; N = 8) dairy cows. The expressions of the acute phase protein genes serum amyloid A3 (SAA3), haptoglobin (HP), ceruloplasmin (CP) genes in the tissues most exposed to pathogens— mammary gland cistern lining epithelial cells (CLECs) and mammary epithelial cells (MECs)—were analyzed.ResultsWe found constitutive and extrahepatic expressions of the studied genes in both tissue types. HP expression in the MECs of the CoPS-infected group was higher than in the H group (p ≤ 0.05). Moreover, higher SAA3 expression in the CoPS and CoNS groups than in the H group (p = 0.06 and 0.08, respectively) was found. No differences between SAA3 and HP in CLECs were revealed, regardless of the pathogen type. However, higher expression of CP (p ≤ 0.05) in the CoPS group than in the H group was noted.ConclusionsThe expressions of selected acute phase proteins were similar between CLECs and MECs, which means that CLECs are not only a mechanical barrier but are also responsible for the biological immune response. Our findings agree with the results of other authors describing the immunological response of MECs during chronic mastitis, but the results for CLECs are novel.

Application and validation of the systemic severity score system for acute coliform mastitis in dairy cattle from farms of various types and sizes

乳牛の急性大腸菌性乳房炎（ACM）の重症例は高率に除籍（死亡あるいは生産性低下により淘汰）されるため重症度評価には迅速性が要求される．全身症状4 項目（体温，第一胃運動性，脱水程度，沈うつ徴候）の合計スコア（1 点間隔・各項目2 〜3 段階；軽症0 〜2，中等症3 〜5，重症6 〜9）により，ACM の重症度を評価するSystemic Severity Score system（SSS 法）は迅速性に優れ，かつ，スコア（SC）と重症度の関連性が検証されている数少ない評価方法の1 つであるが，アメリカの大規模6 農場の調査成績を基に考案されたものであるため，様々な飼養形態や規模の農場を対象とする往診診療における有用性は不明である．そこで，当診療エリアの往診ACM 症例を対象として，飼養状況，初診後100 日以内の除籍について調査し，SSS 法による重症度の妥当性を除籍発生頻度，受信者動作特性曲線（ROC 曲線）分析，生存時間分析を用いて検証した．

O-29 Negative Sexual Outcomes in Patients With Chronic Granulomatous Mastitis from Sao Paulo, Brazil

O-29 Negative Sexual Outcomes in Patients With Chronic Granulomatous Mastitis from Sao Paulo, Brazil

Expression analysis of candidate genes for chronic subclinical mastitis in Norwegian Red cattle

Chronic subclinical mastitis (SCM) is characterized by a long-term inflammation in the udder with high somatic cell count (SCC) in milk. Previously, several novel alternative SCM traits for Norwegian Red (NR) cattle have been defined to improve breeding strategies against chronic SCM. Quantitative trait loci and candidate genes affecting chronic SCM in NR have been identified. The aim of this study was to analyze the expression profiles of 14 selected candidate genes (RAD17, ACOT2, ACOT4, FOS, CXCL1, CXCL8, CCNB1, CDK7, TGFB3, SEL1L, STAT4, C6, GLI2, and SLC18A2). Twenty healthy NR cows with official genomic estimated breeding values (GEBV) for lactation average somatic cell scores (LSCS) were selected. Ten cows had high GEBV for LSCS (cows with low probability to have high SCC in milk during lactation) and 10 cows had low GEBV for LSCS (cows with high probability of having high SCC in milk). We isolated RNA from unstimulated peripheral blood mononuclear cells from these. Two out of the 14 analyzed genes showed significantly different results between groups. The group with high GEBV for LSCS displayed significantly higher expression of the CXCL1 gene than the low GEBV group. Grouping by lactation stage revealed significant differential expression of the FOS gene, with higher expression in early lactation (2-3 mo after calving) compared with late lactation (7-8 mo after calving). In addition, flow cytometry was performed on the peripheral blood mononuclear cells samples to analyze if number and type of isolated cells influenced the gene expression in the groups. The results in the current study provide identified genes that can be considered as possible candidate genes for chronic SCM in NR cows.

THE OCCURRENCE OF BACTERIAL AGENTS CAUSING MASTITIS IN DAIRY SHEEP AND THEIR RESISTANCE TO ANTIBIOTICS

Introduction: The aim of this study was to monitor the occurrence of bacterial agents causing mastitis, forms of mastitis and antibiotic resistance in 300 ewes on a farm in Eastern Slovakia.
 Material and methods: During the milking season, were performed three complex investigations including clinical examination, California Mastitis Test and laboratory analysis of milk samples. The investigations and milk samples were taken in three phases; the beginning (April), the middle (June) and the end (September) of the milking season.
 Results: Of all the samples (806), 225 (28.0%) were positive for bacterial pathogens. The highest incidence of mastitis (33.3%) was recorded in September, while April (23.8%) and June (25.3%) had lower incidence. The samples from September showed the highest incidence of subclinical mastitis (20.1%), with 13.2% being an acute form of mastitis. Coagulase-negative staphylococci (CNS) were identified in 61.7% of the positive samples. Especially, S. chromogenes, S. epidermidis and S. schleiferi were most frequently isolated. Staphylococcus aureus was the causative agent in 20.0% of the positive samples and caused acute or subclinical mastitis in the affected ewes. The tested bacteria showed very high resistance to novobiocin (59.5%) and penicillin (51.4%) and high resistance to amoxycillin (35.1%). We found that 80% of Staphylococcus aureus bacteria tested for antibiotic resistance were resistant to novobiocin and 70% were resistant to penicillin. Of all tested CNS, 56.5% were resistant to novobiocin, 39.1% to penicillin, and 34.7% to amoxicillin.
 Conclusion: Proper isolation and identification of the causative organism play a significant role in the prevention and control of the intramammary infection. In our study, a combinations of Streptomycin, Ciprofloxacin and Tetracycline were the most effective antibiotics for the control of mastitis.

Study of spectrum and clinical profile of benign breast disease in the rural area: is there any change

Background: Recent understanding of pathophysiology and health awareness may have impact on spectrum of benign breast disease (BBD) in rural area. We have analyzed clinical data of 88 cases of BBD for spectrum and clinical profile to compare with present studies.Methods: Clinical records of 88 cases of BBD were reviewed for demographic details, clinical presentation, management and outcome. Recent literature related to BBD in rural area was searched using various search engines. Results of our study were compared with recent studies.Results: Fibroadenoma (FA) was most common condition followed by fibrocystic disease. BBD were common in third decade 46 (52.3%). Lump was present in 87 cases. Lump was painful in 45 (52.3%), and associated with fever in 15 (17.1%) cases. Fine needle aspiration cytology and ultrasound was done in 70 and 32 patients in which diagnostic accuracy with histopathology was seen in 69.7% and 56.2% respectively. Surgical treatment included lumpectomy, lump excision, mastectomy, drainage with debridement and axillary clearance. Three patients of breast tuberculosis were treated with 9 months AKT. When compared with recent studies, over all spectrum was same expect chronic abscess presenting as lump was more in our series.Conclusions: Our study revealed benign neoplasms, inflammatory condition and tumor like lesions in 43.1%, 14.6% and 13.8% cases respectively. FA was the most common lesion. BBD were common in third decade. Except the chronic mastitis, incidence of various types of benign lesions was comparable with those found in other studies. There is not much change in overall spectrum of benign breast disease.

Physiological response of mammary glands to Escherichia coli infection: A conflict between glucose need for milk production and immune response

The mammary immune and physiological responses to distinct mammary-pathogenic E. coli (MPEC) strains were studied. One gland in each of ten cows were challenged intra-mammary and milk composition (lactose, fat, total protein, casein), biochemical (glucose, glucose-6-phosphate (Glu6P), oxalate, malate, lactate, pyruvate and citrate, malate and lactate dehydrogenases, lactate dehydrogenase (LDH), nitrite, lactic peroxidase, catalase, albumin, lactoferrin, immunoglobulin) and clotting parameters were followed for 35 days post-challenge. Challenge lead to clinical acute mastitis, with peak bacterial counts in milk at 16–24 h post-challenge. Biochemical and clotting parameters in milk reported were partially in accord with lipopolysaccharide-induced mastitis, but increased Glu6P and LDH activity and prolonged lactate dehydrogenase and Glu6P/Glu alterations were found. Some alterations measured in milk resolved within days after challenge, while others endured for above one month, regardless of bacterial clearance, and some reflected physiological responses to mastitis such as the balance between aerobic and anaerobic metabolism (citrate to lactate ratios). The results suggest that E. coli mastitis can be divided into two stages: an acute, clinical phase, as an immediate response to bacterial infection in the mammary gland, and a chronic phase, independent of bacteria clearance, in response to tissue damage caused during the acute phase.

Sensitivity and Resistance of the Microbiota of Reproductive Organs and Mammary Gland of Cows to Anti-microbial Agents in Cases of Inflammation

This paper contains the results of complex microbiological studies (culture method, real-time PCR) of biological material from cows affected by inflammatory diseases of reproductive organs and mammary gland. Milk microbiota with underlying subclinical mastitis was represented by pathogenic (S. aureus, enteropathogenic E. coli), opportunistic bacteria (Staphylococcus spp., E. coli, E. faecium, Streptococcus spp., S. agalactiae, P. aeruginosa) and yeast-like fungi of Candida spp. in association. Combined infectious and inflammatory diseases of genital tract and mammary gland in cows resulted in the same types of microorganisms in microflora composition, which confirms information obtained by other researchers on associated microbiota during inflammatory processes. S. aureus, P. aeruginosa, E. coli, E. faecium, E. faecalis, S. epidermidis, S. saprophyticus, Bacillus spp., and yeast-like fungi of Candida spp. were obtained from milk samples from cows with acute clinical mastitis and in vaginal washes from cows with acute postpartum endometritis. In the course of defining the sensitivity to antibiotics and identifying genes of antimicrobial resistance, multidrug resistance was established in most part of obtained strains, which should be taken into account when planning treatment measures.

Acute Phase Protein as Biomarker for Diagnosis of Sub-Clinical Mastitis in Cross-Bred Cows

The present study was carried out to investigate the role of acute phase proteins [Serum amyloid A (SAA) and milk amyloid A (MAA)] for diagnosis of sub-clinical mastitis in cross-bred cows. The study was conducted on 40 apparently healthy cross-bred cows, in the mid-lactation period and were equally divided into four groups [Group-I (Bacteriologically negative milk samples, California mastitis test (CMT) negative), Group-II (Bacteriologically negative with SCC £200,000 cells/ml milk, CMT positive), Group-III (Bacteriologically negative with SCC ³200,000 £500,000 cells/ml milk, CMT positive) and Group-IV (Bacteriologically positive with SCC ³200,000 £500,000 cells/ml milk, CMT positive)]. On bacteriological examination of 57 milk samples, 27(47.36%) were found positive and 30 (52.63%) were found negative. A highly significant (P 200,000 cells/ml, 74 µg/ml and 10 µg/ml, respectively for the diagnosis of sub-clinical mastitis with high sensitivity (90% to 100%). The area under curve of the SAA was larger than those of SCC and MAA, which suggests that the SAA test was more accurate than SCC and MAA for the diagnosis of sub-clinical mastitis.

Gangrenous mastitis in dromedary camels in UAE caused by Streptococcus agalactiae

BackgroundMastitis is a disease of economic concern that affects dairy industry worldwide. This study aimed to investigate and identify possible etiologies encountered in an episode of acute gangrenous mastitis in lactating she-camels in Al Dhafra region, Abu Dhabi Emirate, United Arab Emirates (UAE).Beside the routine clinical examination, conventional bacteriological methods were used to isolate and identify possible aerobic/anaerobic bacterial or fungal pathogens from cultured milk samples collected from the mastitic she-camels. Moreover, quantitative real-time polymerase chain reaction (qPCR) was used for the detection of Mycoplasma agalactiae and Mycoplasma bovis strains, and the 16S rRNA gene was sequenced to confirm the isolation. The isolates were also tested for their susceptibility to antimicrobials.ResultsAcute gangrenous mastitis is reported in the dromedary camel herd with about 80% morbidity rate among lactating she-camels exhibited acute, painful hard swelling of affected teat, quarter or entire udder. About 41.7% of the infected animals were stamped out for culling due to complete or partial amputation of udder quarters. Streptococcus agalactiae was the sole isolated organism (6 isolates). The antimicrobial susceptibility testing revealed that, the Streptococcus agalactiae isolates were sensitive to both penicillin and ampicillin. Comparison of the 16S rRNA gene sequencing results by BLASTN confirmed the presence of Streptococcus agalactiae with high confidence (100% identity). Phylogenetic analysis indicated clustering of one isolate (CMAUAE accession number; MN267805.1) with Streptococcus agalactiae that infects multi-hosts including humans, while strains (CMBUAE to CMFUAE with accession numbers; MN267806.1 to MN267810.1 respectively) clustered with Streptococcus agalactiae that infects humans. No Mycoplasma spp was detected by qPCR analysis.ConclusionsIn the present study, the Streptococcus agalactiae was found to be the main cause of acute gangrenous mastitis in dromedary camels in UAE. More research should be done to investigate other possible causes of clinical or subclinical mastitis in dromedary camels in UAE.

Some Udder Problems associated with productivity in Goats

The aim of this article was study the mastitis of a goat under farmer Egyptian conditions. This work was conducted on a private farm containing50baladigoats suffering from symptoms of mastitis. All environmental factors were recorded as housing condition, breeding, health problems, production and feeding. Morphological examination of the udder were carried out. Milk samples of infected goats, were collected for bacterial isolation and sensitivity test. Animals were treated with systemic and local intramammary broad spectrum antibiotic. The results indicated that, 35 cases (70 %) were responded to treatment, and showed normal reproductive and productive performances.15 cases (30 %) of chronic mastitis do not respond to treatment and breeding. Post-mortem examination of these cases revealed widespread diffused chronic mastitis throughout the affected udder. Tissue specimens of infected udder and supra mammary lymph node were fixed in 10% formalin for histopathological examination. Bacteriological examination revealed isolation of Staph aureus, Corynebacterium pyogenes, Salmonella spp., E coli in a single or mixed with other bacteria as Pasteurella spp. Fungus suspect Asperigillus spp. infection and yeast, suspect Candida spp. Pathological examination revealed chronic diffused suppurative and chronic granulomatous mycotic mastitis. In conclusion, mastitis is an important disease of goats and prevention of infection is the key for disease control. Good environ mental and hygienic housing of milking practices are a necessity to minimize the impact of this disease.

Efficacy and safety of oncoplastic surgery plus drug therapy for chronic tuberculous granulomatous mastitis.

BACKGROUND: Chronic tuberculous granulomatous mastitis (CTGM) is a rare form of tuberculosis (TB) treated primarily with anti-TB drugs. Oncoplastic surgery (OS) has been proposed as adjuvant therapy for CTGM.METHOD: We followed for 1 year every CTGM patients and assessed the efficacy (defined as non-recurrence and no need for corticosteroids) and safety attributable to the standard anti-TB drugs therapy with and without OS.RESULTS: We analysed 128 CTGM cases, including 78 (61%) treated with OS plus anti-TB drugs and 50 (39%) with anti-TB drugs only. We observed a significantly higher efficacy among those exposed vs. unexposed to OS (100% vs. 92%; prevalence ratio [PR] 1.09, 95% CI 1.00-1.18), with no difference in the number of complications (21% vs. 8%; PR 2.56, 95% CI 0.91-7.26). We also observed that the incidence of post-operative complications decreased by 50% when OS was postponed from after Month 1 to after completing Month 2 of anti-TB drugs treatment (19% to 8%; PR 0.46, 95% CI 0.13-1.62).CONCLUSION: OS appears to represent an efficacious and safe adjuvant therapy when combined with anti-TB drugs in the treatment of CTGM patients, but clinical trials are needed to prove this observation.

Identification of a reliable fixative solution to preserve the complex architecture of bacterial biofilms for scanning electron microscopy evaluation.

Bacterial biofilms are organized sessile communities of bacteria enclosed in extracellular polymeric substances (EPS). To analyze organization of bacteria and EPS in high resolution and high magnification by scanning electron microscopy (SEM), it is important to preserve the complex architecture of biofilms. Therefore, fixation abilities of formalin, glutaraldehyde, and Methacarn (methanol/chloroform/acetic acid-6:3:1) fixatives were evaluated to identify which fixative would best preserve the complex structure of bacterial biofilms. Economically important Gram-negative Mannheimia haemolytica, the major pathogen associated with bovine respiratory disease complex, and Gram-positive Staphylococcus aureus, the major cause of chronic mastitis in cattle, bacteria were selected since both form biofilms on solid-liquid interface. For SEM analysis, round glass coverslips were placed into the wells of 24-well plates and diluted M. haemolytica or S. aureus cultures were added, and incubated at 37°C for 48-72 h under static growth conditions. Culture media were aspirated and biofilms were fixed with an individual fixative for 48 h. SEM examination revealed that all three fixatives were effective preserving the bacterial cell morphology, however only Methacarn fixative could consistently preserve the complex structure of biofilms. EPS layers were clearly visible on the top, in the middle, and in the bottom of the biofilms with Methacarn fixative. Biomass and three-dimensional structure of the biofilms were further confirmed spectrophotometrically following crystal violet staining and by confocal microscopy after viability staining. These findings demonstrate that Methacarn fixative solution is superior to the other fixatives evaluated to preserve the complex architecture of biofilms grown on glass coverslips for SEM evaluation.

Pathogen effects on milk yield and composition in chronic subclinical mastitis in dairy cows

This study aimed to evaluate the effects of chronic subclinical mastitis (CSM) on milk production and component yields in dairy cows. A total of six herds located in the Midwest area of São Paulo State, Brazil were selected. Herds were visited once every 2 weeks to measure milk yield and to collect milk samples from lactating Holstein cows. Milk samples were collected at two stages (1 and 2), and each stage comprised three milk samplings. In stage 1, a total of 117 of 647 cows were diagnosed with CSM based on at least two of three repeated somatic cell counts (SCC) > 2000,000 cells/mL and positive bacterial milk culture results (BC). Cows with CSM were selected for the second stage. In stage 2, selected cows had quarter sampling aseptically collected for BC analyses prior to milking, and quarter milk yield was measured. Milk components (total protein, fat, lactose, and total solids) were measured using mid-infrared spectroscopy. Mammary quarters were considered healthy if all three repeated SCC results were ≤ 200,000 cells/mL and no bacterial growth was detected on BC. All quarters with positive bacterial growth were classified as having (non-chronic) subclinical mastitis when only one of three SCC results were > 200,000 cells/mL, and CSM when at least two of three SCC results were > 200,000 cells/mL. The effects of CSM by type of pathogen on milk and components yield were assessed using a linear mixed model.Mammary quarters with CSM caused by major pathogens had milk loss of 1.1 kg/quarter milking in comparison to healthy quarters. Milk losses were 0.8 and 1.3 kg/quarter milking when CSM was caused by Staphylococcus aureus or environmental streptococci, respectively. In addition, healthy quarters produced more milk components than quarters with CSM caused by major pathogens. Minor pathogens causing CSM (non-aureus staphylococci and Corynebacterium spp.) had no effect on milk yield. Quarters with CSM had lower milk and component yields when compared with healthy quarters. Milk losses varied according to the type of pathogen and were higher when associated with major pathogens such as S. aureus and environmental streptococci compared with healthy quarters.

Chronic subclinical mastitis reduces milk and components yield at the cow level.

We evaluated the effects of chronic subclinical mastitis (CSM) caused by different types of pathogens on milk yield and milk components at the cow level. A total of 388 Holstein cows had milk yield measured and were milk sampled three times at intervals of two weeks for determination of SCC and milk composition, and microbiological culture was performed. Cows were considered healthy if all three samples of SCC were ≤200 000 cells/ml and were culture-negative at the third milk sampling. Cows with one result of SCC > 200 000 cells/ml were considered to suffer non-chronic subclinical mastitis whereas cows with at least 2 out of 3 results of SCC > 200 000 cells/ml had CSM. These latter cows were further sorted according to culture results into chronic negative-culture or chronic positive-culture. This resulted in four udder health statuses: healthy, non-chronic, chronicNC or chronicPC. The milk and components yields were evaluated according to the udder health status and by pathogen using a linear mixed effects model. A total of 134 out of 388 cows (34.5%) were chronicPC, 57 cows (14.7%) were chronicNC, 78 cows (20.1%) were non-chronic and 119 cows (30.7%) were considered healthy, which resulted in a grand total of 1164 cow records included in the statistical model. The healthy cows produced more milk than each of the other groups (+2.1 to +5.7 kg/cow/day) and produced higher milk component yields than the chronicPC cows. The healthy cows produced more milk than cows with chronicPC caused by minor (+5.2 kg/cow/day) and major pathogens (+7.1 kg/cow/day) and losses varied from 5.8 to 11.8 kg/cow/day depending on the pathogen causing chronicPC mastitis. Chronic positive-culture cows had a reduction of at least 24.5% of milk yield and 22.4% of total solids yield.

Antimicrobial susceptibility of nine udder pathogens recovered from bovine clinical mastitis milk in Europe 2015–2016: VetPath results

VetPath is an ongoing pan-European antimicrobial susceptibility monitoring programme collecting pathogens from diseased cattle, pigs and poultry not recently treated with antibiotics. Non-duplicate isolates (n = 1244) were obtained from cows with acute clinical mastitis in eight countries during 2015–2016 for centrally antimicrobial susceptibility testing according CLSI standards.Among Escherichia coli (n = 225), resistance was high to ampicillin and tetracycline, moderate to kanamycin and low to amoxicillin/clavulanic acid and cefazolin. The MIC50/90 of danofloxacin, enrofloxacin and marbofloxacin were 0.03 and 0.06 μg/mL. For Klebsiella spp. (n = 70), similar results were noted, except for ampicillin and kanamycin. We detected 3.7 % (11/295) Enterobacteriaceae isolates carrying an ESBL/AmpC gene. Staphylococcus aureus (n = 247) and coagulase-negative staphylococci (CoNS; n = 189) isolates were susceptible to most antimicrobials tested except to penicillin (25.1 and 29.1 % resistance). Two S. aureus and thirteen CoNS isolates harboured mecA gene. Streptococcus uberis isolates (n = 208) were susceptible to β-lactam antibiotics (87.1–94.7 % susceptibility), 23.9 % were resistant to erythromycin and 37.5 % to tetracycline. Resistance to pirlimycin was moderate. For Streptococcus dysgalactiae (n = 132) the latter figures were 10.6 and 43.2 %; pirlimycin resistance was low. MIC values for Streptococcus agalactiae, Trueperella pyogenes and Corynebacterium spp. were generally low.This current VetPath study shows that mastitis pathogens were susceptible to most antimicrobials with exceptions of staphylococci against penicillin and streptococci against erythromycin or tetracycline. For most antimicrobials, the percentage resistance and MIC50/90 values among the major pathogens were comparable to that of the preceeding VetPath surveys. This work highlights the high need to set additional clinical breakpoints for antimicrobials frequently used to treat mastitis.

Hepatic Transcriptome Analysis Identifies Divergent Pathogen-Specific Targeting-Strategies to Modulate the Innate Immune System in Response to Intramammary Infection.

Mastitis is one of the major risks for public health and animal welfare in the dairy industry. Two of the most important pathogens to cause mastitis in dairy cattle are Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). While S. aureus generally induces a chronic and subclinical mastitis, E. coli is an important etiological pathogen resulting in an acute and clinical mastitis. The liver plays a central role in both, the metabolic and inflammatory physiology of the dairy cow, which is particularly challenged in the early lactation due to high metabolic and immunological demands. In the current study, we challenged the mammary glands of Holstein cows with S. aureus or E. coli, respectively, mimicking an early lactation infection. We compared the animals' liver transcriptomes with those of untreated controls to investigate the hepatic response of the individuals. Both, S. aureus and E. coli elicited systemic effects on the host after intramammary challenge and seemed to use pathogen-specific targeting strategies to bypass the innate immune system. The most striking result of our study is that we demonstrate for the first time that S. aureus intramammary challenge causes an immune response beyond the original local site of the mastitis. We found that in the peripheral liver tissue defined biological pathways are switched on in a coordinated manner to balance the immune response in the entire organism. TGFB1 signaling plays a crucial role in this context. Important pathways involving actin and integrin, key components of the cytoskeleton, were downregulated in the liver of S. aureus infected cows. In the hepatic transcriptome of E. coli infected cows, important components of the complement system were significantly lower expressed compared to the control cows. Notably, while S. aureus inhibits the cell signaling by Rho GTPases in the liver, E. coli switches the complement system off. Also, metabolic hepatic pathways (e.g., lipid metabolism) are affected after mammary gland challenge, demonstrating that the liver restricts metabolic tasks in favor of the predominant immune response after infection. Our results provide new insights for the infection-induced modifications of the dairy cow's hepatic transcriptome following mastitis.

Role of Artesunate in Potentiation of β-lactam Against Methicillin Resistant Staphylococcus aureus (MRSA) Isolated from Bovine Mastitis and its Histopathology Impact In-Vivo Study

This article discusses the augmenting influence of Artesunate (ART) in combination with β-lactams (amoxicillin/clavulanic acid) antibiotic in sepsis mice models infected by a lethal challenge dose of live coagulase positive enterotoxigenic (Sec) MRSA that was isolated from a case of chronic bovine mastitis. The main goal is to find an appropriate treatment to overcome resistance mechanism of MRSA towards β-lactams antibiotic. Fifty healthy adult Swiss mice divided into 5 equal groups were used in the experimental procedure. The infected group that treated with both ART and β-lactams (amoxicillin/clavulanic acid) antibiotic revealed complete inhibition of MRSA count with complete normal macroscopic and histopathological features. We suggest that ART can potentiate the antibacterial action of β-lactams (amoxicillin/Clavulanic) acid against MRSA infection. The combination of ART and antibiotic can overcome MRSA resistance mechanism and so could be considered a novel candidate to overcome mastitis and/or sepsis caused by MRSA.

Spectrum of Breast Lesions on Fine Needle Aspiration Cytology in Punjab

<p><strong>Background and Objectives: </strong>Breast lumps can be diagnosed in a short time by fine needle aspiration cytology (FNAC). Moreover, being cost effective, it is rendered the first choice by surgeons and patients.This study was carried out to outline the spectrum of breast lesions on FNAC in patients presenting with breast lumps in two tertiary care teaching hospitals of Punjab.<br /><strong>Methods:</strong> A cross sectional descriptive study was conducted from January, 2018 to October, 2019.<br />Hundred cases of lumps in breast were taken from Sahara Medical College and Sughra Shafi Medical Complex, Narowal and Lahore General Hospital, Lahore. Cytological diagnosis was made and correlated clinically and histologically. Data analysis was done by Statistical Package for the Social Sciences (SPSS) version 24.0 and expressed as frequency and percentages.<br /><strong>Results:</strong> Majority (58%) of cases were Class-2 smears on cytology while 11% were malignant and<br />belonged to Class-5. Among the benign lumps, fibroadenoma was the most common (55.1%) followed by fibrocystic disease (17.2%) and acute mastitis (12%). Thirty-one cases were in grey zone i.e. Class-3 smears showing atypia and Class-4 smears that were suspicious of malignancy.<br /><strong>Conclusion:</strong> By screening patients with lumps in breast, early and effective management can be made possible. Class 3 and 4 smears should be confirmed with excision and biopsy of the lump for conclusive diagnosis.</p>

Serum iron concentration in cattle with endotoxaemia.

The objective of this study was to examine whether serum iron (Fe) concentration is useful as a prognostic biomarker for cows with acute coliform mastitis (ACM). Our study was composed of determining the reproducibility of serum Fe concentration as a prognostic criterion in cows with ACM (Study 1) and clarifying the sequential changes in serum Fe concentration in cattle that received endotoxin (Study 2). Seventy-seven cows with (n = 47) or without (n = 30) ACM were enrolled in Study 1. The proposed diagnostic cut-off value of serum Fe concentration indicating a poor prognosis of ACM based on the analysis of the receiver operating characteristic curves was < 31.5 µg/dL. Ten young cattle aged 176.8 ± 23.7 days were enrolled in Study 2. Five young cattle received endotoxin (LPS group) and the remaining five received physiological saline (control group). Blood collections were carried out before endotoxin challenge (pre), and 0.5, 1, 2, 4, 8, 12, 24, and 48 h after the challenge. As a result, a significant decrease in serum Fe concentration was not observed until 24 h after endotoxin challenge. Because in cows with clinical ACM it is difficult to know the time course after infection, the alteration in serum Fe concentrations alone may be an insufficient prognostic criterion.