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Contents This study compared endometrial cytology vis-a-vis uterine fluid cytology for assessment of uterine health in clinically normal and subclinical endometritis (SE)-affected buffaloes. Uterine fluid samples and endometrial samples were collected from the buffaloes ( n = 38) at oestrus using blue sheath and cytobrush, respectively. The smears were stained with Field stain for 3 minutes, and a minimum of 400 cells were counted in each smear for determination of the percentage of polymorphonuclear (PMN) leucocyte. The incidence of subclinical endometritis, based on the cytobrush cytology, was 23.08%. The correlation between cytobrush cytology with uterine fluid cytology was positive and significant ( r = .37; p = .02). The ratio of PMN leucocyte in cytobrush cytology to uterine fluid cytology was 1:2.4. ROC analysis revealed that the threshold value of 6.16% PMN leucocyte in uterine fluid cytology showed a diagnostic sensitivity and specificity of 100% in differentiating normal from SE-affected buffaloes. In conclusion, collection of uterine fluid was easier compared to collection of endometrial samples using cytobrush and the percentage of PMN leucocyte in uterine fluid cytology can be used as a tool for diagnosis of subclinical endometritis in buffaloes. [ABSTRACT FROM AUTHOR]

Background: Buffalo reactivity during milking affects milking procedures, milk yield, and quality., Aims: This study evaluated the influence of milking reactivity on the yield, composition, somatic cell count, pH, and electrical resistance of milk in Jaffarabadi buffaloes., Methods: A 1-4 point scale, based on leg movement, was used to assess the milking reactivity of buffaloes (n=40). Based on the milking reactivity score, animals were classified into four groups: reactivity score-1 (RS1), reactivity score-2 (RS2), reactivity score-3 (RS3), and reactivity score-4 (RS4). The influence of milking reactivity on yield, composition, somatic cell count, pH, and electrical resistance of milk was observed., Results: Buffaloes with RS1 and RS2 produced significantly (P≤0.001) higher daily milk yield, 6% fat-corrected yield, solid-corrected yield, and energy-corrected yield than the RS3+4 group. Milking reactivity score did not influence milk fat, protein, lactose, ash, solid-not-fat, total solids content, and the fat: protein ratio. However, daily yield of milk fat (P<0.001), protein (P=0.001), lactose (P=0.001), ash (P=0.002), solid-not-fat (P=0.001), and total solids (P<0.001) were significantly higher in buffaloes in the RS1 and RS2 groups than in the RS3+4 group. Milk somatic cell count and somatic cell score were not influenced by milking reactivity score (P>0.05). Milk density and pH did not differ significantly (P>0.05) between groups. However, the electrical resistance of milk in the RS1 group was significantly (P<0.05) higher than in the RS2 and RS3+4 groups., Conclusion: Milking reactivity influences daily milk yield, milk component yield, and electrical resistance, but not milk composition in Jaffarabadi buffaloes., Competing Interests: The authors declare that there was no conflict of interest.

Background: The photoperiod and other seasonal variations are the key factors that affect reproduction and production of the animals. The pineal gland secretes melatonin hormone that affects several physiological functions of the body during different seasons., Aims: The present study was conducted to study the histoarchitectural and micrometrical changes in the pineal gland of buffalo ( Bubalus bubalis ) during different seasons of the year., Methods: Pineal glands of 30 adult female Jaffarabadi buffaloes were collected from the slaughterhouse during the winter, summer, and rainy seasons. Samples were processed by standard histological procedures and stained with various stains for histological and micrometrical observations., Results: The pinealocytes constituted a major cellular portion of pineal parenchyma. The pinealocyte nuclei were lightly stained and more euchromatic during the winter season whereas darkly stained and slightly heterochromatic during summer. The calcium deposits occupied a larger area of pineal parenchyma during the summer as compared to the winter season. The pinealocyte density, the nuclear diameter of pinealocytes, and the number of argyrophilic nucleolar organizer regions (AgNOR) were highest during the winter season as compared to the summer and rainy seasons., Conclusion: The present study shows the influence of season on the histoarchitecture and histometry of the pineal gland of buffalo and indicated higher pineal activity during the winter season in this species., Competing Interests: The authors declare that there is no conflict of interest.

The success of assisted reproduction relies on functional competence of frozen-thawed semen. Heat stress affects protein folding leading to aggregation of mis-folded proteins. Hence, a total of 384 (32 ejaculates/bull/season) ejaculates from six matured Gir bulls were used to evaluate physico-morphological parameters, the expression of HSPs (70 and 90) and fertility of frozen-thawed semen. The mean percent individual motility, viability and membrane integrity were significantly ( p <  0.01) higher in winter compared to summer. Out of 1200 Gir cows inseminated, 626 confirmed pregnant and the mean conception rate of winter (55.04 ± 0.35) was significantly ( p <  0.001) higher than summer (49.33 ± 0.32). A significant ( p <  0.01) difference in concentration of HSP70 (ng/mg of protein) but not HSP90was observed between the two seasons. The HSP70 expression in pre-freeze semen of Gir bulls had significant positive correlation with motility ( p <  0.01, r  = 0.463), viability ( p <  0.01, r  = 0.565), acrosome integrity ( p <  0.05, r  = 0.330) and conception rate ( p <  0.01, r  = 0.431). In conclusion, the season influences physico-morphological parameters and expression of HSP70 but not HSP90 in Gir bull semen. The HSP70 expression is positively correlated with motility, viability, acrosome integrity and fertility of semen. The semen expression of HSP70 may be utilized as biomarker for thermo-tolerance, semen quality and fertilizing capacity of Gir bull semen.

Ethnopharmacological Relevance: A combination of Trachyspermum ammi L., Curcuma longa L., Cuminum cyminum L., Trigonella foenum-graecum L., Foeniculum vulgare Mill., Anethum graveolens L and Zingiber officinale Roscoe is used as immunity booster and reproductive efficiency enhancing agents in folklore medicine., Aim of the Study: The present study aimed to assess the immunomodulatory, uterine cleansing and reproduction enhancing effects of polyherbal mixture in post-partum buffaloes., Materials and Methods: Enzyme linked immunosorbent assay (ELISA) was used to investigate the effects of polyherbal mixture feeding on for quantification of neutrophil functions and blood progesterone hormone estimation. Ultrasonography was used to assess the status of uterine involution, fluid in uterus and ovarian follicular status. Quantitative real time PCR (qRT-PCR) was used to measure the expression of chemokine genes CXCR1, CXCR2 AND IL-8. Artificial insemination with cryopreserved semen was used to breed the animals. Reproductive efficiency parameters were assessed using standard calculation methods., Results: Neutrophil functions and transcriptional abundance of chemokine genes were significantly (P < 0.05) higher in buffaloes supplemented with polyherbal mixture compared to buffaloes in control group. The rate of cervical and uterine involution was significantly (P < 0.05) higher in treatment group compared to control group. The service period was shorter, days to first insemination was earlier and the number of services per conception was lower in buffaloes supplemented with polyherbal mixture compared to the buffaloes in control group. The proportion of buffaloes with large ovarian follicles within 28 days of post-partum was also significantly (P < 0.05) higher in treatment group compared to the control group., Conclusions: The polyherbal mixture used in the study improved the immunity of the buffaloes, facilitated early involution of cervix and uterus, efficient cleansing of lochia and improved subsequent fertility. It has the potential to be used in dairy animals for improving post-partum reproductive efficiency., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A total of 1455 local and non-local (originating from other Indian states), slaughtered or spontaneously dead, sheep in various areas of Kashmir Valley were investigated for the presence of cystic echinococcosis over a period of one year. The overall prevalence was 7.97% with higher prevalence in local (14.3%) than in non-local sheep (6.06%). The prevalence of infection, total number of cysts recovered and mean intensity of infection were higher in lungs as 66.2%, 506 & 5.1% respectively, followed by liver (28.5%, 169, 3.9%) and spleen (5.3%, 9, 1.13%). Either single (71.55%) or multiple (28.45%) organ involvements were observed. 66.6% of cysts were of small size, 19.29% medium, 7.01% large and 7.01% calcified. The fertility of cysts was noted to be 65.7% whereas 34.2% were infertile which included 27.1% sterile and 7.01% calcified cysts. The viability percentage of protoscolices from all the fertile cysts was 74.2%. The number of cysts recovered was higher in sheep with body condition score- emaciated, thin and average, and lower in, fat and obesed. The study showed that the local sheep were more vulnerable to contract cystic echinococcosis than non-local sheep which is further aggravated by poor body condition., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Oxidative stress during peripartum period may compromise the uterine immunity. In the present study, we assessed the oxidative stress and antioxidant status during peripartum period and studied their relationship with postpartum uterine infection in dairy cows. Peripheral blood concentrations of total antioxidant capacity (TAC), malondialdehyde (MDA) and nitric oxide (NO) were determined (day -21, -7, on the day of calving and day +7, +21, +35) in normal (n=11), puerperal metritic (n=7) and clinical endometritic (n=6) cows. Endometrial biopsy was performed on the day of calving and expression of CAT, GPx4 and SOD2 genes was studied using qRT-PCR. Puerperal metritic cows had significantly (P<0.05) lower TAC (on day -7, day 0, day +7, +21 & +35), higher MDA (on day -21, -7 & on the day of calving) and NO (on day 0, +7 & day +35) concentrations compared to normal cows. Similarly, clinical endometritic cows had significantly (P<0.05) lower TAC (on day -7, 0, +7 & +21), higher MDA (on day -21, -7, +7 and +35) and NO (on day +7, +21 & +35) concentrations compared to normal cows. The expression of CAT and GPx4 genes was lower (P<0.05) and SOD2 gene was higher (P<0.05) in endometrial tissue of cows that developed uterine infection compared to normal cows. The relationship of peripheral levels of MDA and NO with antioxidant enzymes expression in endometrial tissue was found significant. Receiver operator characteristic analysis revealed that the concentrations of TAC on day -7 to day +35, MDA on day -21 to day +7 and NO on the day of calving to day +35 were highly correlated to the development of postpartum uterine infection in cows. It may be inferred that the low serum TAC level and high level of lipid peroxidation and NO during peripartum period influenced the endometrial expression of anitioxidative genes that compromised the uterine health during postpartum period., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Coprological examination of 416 bovine faecal samples revealed the presence of parasitic stages of Toxocara vitulorum , strongyles, Strongyloides spp., Fasciola spp., amphistomes, coccidia ( Eimeria spp. and Cryptosporidium spp.) and Buxtonella sulcata . About 42 % (n = 302) faecal samples from cattle and 36 % (n = 114) samples from buffaloes were positive for gastrointestinal (GI) parasitic infections. Both cattle (14.57 %) and buffalo (15.79 %) had the highest incidence of Buxtonella sulcata , respectively. The overall incidence of GI parasitic infections in young animals (below 1 year) was higher followed by older (more than 5 years) and adult animals (1-5 years) and the difference was statistically significant ( p  < 0.05). Non descriptive breeds of bovines showed more parasitic infections than pure breeds, the difference being statistically non-significant (p > 0.05). Season wise GI parasitic infections were recorded to be non-significantly (p > 0.05) higher in monsoon (48.38 %) followed by summer (39 %) and winter (34.61 %) in cattle. There was no significant variation of GI infections in buffaloes in relation to season though highest prevalence was documented in monsoon (44.89 %) followed by winter (35.71 %) and summer (24.32 %). Similarly, sex wise females recorded higher infection rates than males in bovines and the difference being statistically non-significant (p > 0.05).

Aim: The objective of this study was to investigate the ability of infrared thermography (IRT) technique and its interrelationship with conventional mastitis indicators for the early detection of mastitis in Holstein Friesian (HF) crossbred cows., Materials and Methods: A total of 76 quarters of lactating HF crossbred ( Bos indicus × Bos taurus ) cows (n=19) were monitored for body temperature (i.e., eye temperature) and udder skin surface temperature (USST) before milking using forward-looking infrared (FLIR) i5 camera. Milk samples were collected from each quarter and screened for mastitis using Somatic Cell Count (SCC), Electrical Conductivity (EC), and California mastitis test. Thermographic images were analyzed using FLIR Quick Report 1.2 image analysis software. Data on body and USST were compiled and analyzed statistically using SPSS 16.0 and Sigmaplot 11., Results: The mean±standard deviation (SD) body (37.23±0.08°C) and USST (37.22±0.04°C) of non-mastitic cow did not differ significantly; however, the mean USST of the mastitis-affected quarters were significantly higher than the body temperature and USST of unaffected quarters (p<0.001). The mean±SD USST of the subclinical mastitis (SCM) and clinical mastitis-affected quarters were 38.08±0.17 °C and 38.25±0.33 °C, respectively, which is 0.72 and 1.05 °C higher than the USST temperature of unaffected quarters. The USST was positively correlated with EC (r=0.95) and SCC (r=0.93). The receiver operating characteristic curve analysis revealed a higher sensitivity for USST in early prediction of SCM with a cut-off value of >37.61°C., Conclusion: It is concluded that infrared thermal imaging technique could be used as a potential noninvasive, quick cow-side diagnostic technique for screening and early detection of SCM and clinical mastitis in crossbred cows.

Aim: Present study aimed to evaluate the different episodes of clinical mastitis (CM) and influence of duration of treatment and seasonality on the occurrence of different episodes of CM in crossbred cows., Materials and Methods: A total of 1194 lactation data of crossbred CM cows were collected from mastitis treatment record from 2002 to 2012. Data of CM cows were classified into types of episodes (pattern of repeated or multiple episodes occurrence) and number of episodes (magnitude of multiple cases). Types of episodes were divided as single (clinical cure by a single episode of treatment), relapse (retreatment of the same cow within 21 days), recurrence (new CM at least 21 days after treatment), and both (relapse and recurrence). The season was classified as winter (December to March), summer (April to June), rainy (July to September), and autumn (October to November). The difference between incidences of different types of CM episodes and the association between number or type of CM episodes with duration of treatment and seasons of CM occurrence were analyzed by Chi-square test., Results: Among 1194 animals suffered with CM, 53, 16, and 18% had the single episode, relapse, and recurrence, respectively; while 13% suffered by both relapse and recurrence. We estimated the duration of treatment and found 80% of the cows treated 1-8 days, in which 65% treated for 1-4 days, while 35% cows were treated for 5-8 days. Further, 12% cows treated for 9-15 days and 7.5% cows treated >15 days. The relationship between duration of treatment and different episodes of CM revealed that 1-8 days treated cows were mostly cured by the single episode with less relapse and recurrence. In contrast, the incidences of recurrence and relapse episodes were higher in cows treated for more than 9 days. The highest incidence of relapse was noticed in winter (36%) than other seasons (10-28%), while the recurrence was less during autumn (9%) compared to other seasons (20-40%)., Conclusion: Cows those suffered by both relapse and recurrence were more susceptible to CM, and they need to be culled from farm to control the transmission of infections. Although the influence of seasonality was difficult to understand, the higher magnitude of relapse and recurrence during winter suggested the adverse effects of cold stress on treatment outcome.

Aim: To study the alteration of major milk components such as milk fat, protein, lactose, solid not fat (SNF) and total solids (TS) and their association with different degree of intra-mammary inflammation (IMI) in Jaffrabadi buffaloes., Materials and Methods: Milk samples (n=1516) were collected from Jaffrabadi buffaloes separately from each quarter. Milk samples were analyzed for milk fat, protein, lactose, SNF and TS percent on the same day using milk analyzer "LACTOSCAN." Milk samples were checked for IMI by California mastitis test (CMT), and the results were expressed as negative (0), +, ++, and +++ CMT score. The traits of milk components which showed significant difference (p<0.05) between samples from inflamed and non-inflamed quarters were analyzed by receiver operating characteristic (ROC) analysis to see the accuracy and degree of association with IMI., Results: Among several milk components, milk protein and lactose percent showed a significant difference (p<0.05) between milk samples from normal and inflamed quarters. Though, during the early stage of mammary gland inflammation milk protein percent remained significantly high (p<0.05), later with an increase in the degree of severity of inflammation it did not show any difference. Milk samples from normal udder quarters had significantly higher lactose percent than inflamed quarters (p<0.05). Milk lactose percent decreased gradually with an increase in the degree of severity of inflammation. ROC analysis revealed that milk samples having lactose content below the threshold values had significantly higher chances to come from inflamed udder quarters (p<0.05). Though, the value of the area under curve (AUC) indicated that milk lactose was significantly associated with IMI (p<0.05), the accuracy was moderate (AUC=0.71-0.75)., Conclusions: The results of the present study indicated that milk lactose percent gradually and significantly reduced during IMI and can be used as a marker for identification of IMI in buffaloes. However, ROC analysis further confirmed that using milk lactose IMI can be identified with moderate accuracy.

Retention of fetal membranes (RFM) adversely affects the production and reproduction potential of the affected cows leading to huge economic loss. Physiological separation of fetal membranes is reported to be an inflammatory process. The present study compared the concentrations of certain pro inflammatory cytokines [Interleukin 1β (IL-1), Interleukin 6 (IL-6), Interleukin 8 (IL-8) and Tumor necrosis factor α (TNF-α) between the cows that developed RFM (n=10) and the cows that expelled fetal membranes normally (n=10) to find out if they could serve as a predictive tool for RFM. Blood samples were collected from the cows from 30 days before expected parturition through day -21, day -14, day -7, day -5, day -3, day -1, on the day of parturition (day 0), day 1 postpartum and the pro-inflammatory cytokines were estimated in blood plasma by ELISA method. The IL-1β concentration was significantly lower (P<0.05) in cows that developed RFM compared to those that expelled fetal membranes normally from 3 days before calving till the day of calving. The plasma concentrations of IL-6 and IL-8 were also lower (P<0.05) in cows that developed RFM than those calved normally. On the day of calving, significantly (P<0.05) lower concentrations of TNF-α was observed in cows that developed RFM compared to those expelled fetal membranes normally. It may be inferred that the concentrations of IL-1, IL-6, IL-8 and TNF-α around parturition were altered in cows developing RFM compared to those expelled fetal membranes normally., (Copyright © 2015. Published by Elsevier B.V.)

The risk factors and impact of retained fetal membranes (RFM) on productive and reproductive performance of crossbred cattle, Zebu cattle, and Murrah buffalos were evaluated using data spread over 12 years. Multivariable logistic regression model was used to identify risk factors and to quantify their odds ratio (OR). Overall incidence of RFM in crossbred cattle, Zebu cattle, and Murrah buffalos were 26, 16, and 13 %, respectively; and significant risk factors for RFM in crossbred cattle were abortion (OR = 3.9), dead calf (OR = 4.1), dystocia (OR = 4.3), pluriparity (OR = 1.5), and shorter gestation length (OR = 4.3). In Zebu cattle, abortion (OR = 4.0), dead calf (OR = 3.7), dystocia (OR = 3.9), lower birth weight of calf (OR = 1.6), and shorter gestation length (OR = 6.4) were significant risk factors for RFM. In Murrah buffalos, abortion (OR = 19.2), dead calf (OR = 4.4), dystocia (OR = 4.7), pluriparity (OR = 1.7), shorter gestation length (OR = 12.7), and calving during summer season (OR = 1.8) were the risk factors for RFM. Although the occurrence of RFM did not affect fertility parameters, a significant (P < 0.05) decrease in 305-day milk yield and total milk yield was observed in RFM-affected crossbred cattle. Taken together, it may be concluded that increased parity, abnormal calving, and short gestation length were the main risk factors for RFM in dairy bovine.

Neutrophils employ both oxidative and non oxidative mechanisms to destroy pathogens. Function of neutrophils coming in milk during mammary invasion is not clearly understood in dairy animals. Therefore, the present study was designed in 36 Sahiwal cows to see the changes occurring in the neutrophil activity of cows suffering from subclinical (SCM) and clinical mastitis (CM). Cows were divided into three groups as healthy (n = 12), SCM (n = 12) and CM (n = 12) groups on the basis of CMT scoring, gross morphological changes in milk, bacteriological examination of milk and by counting milk SCC. Significantly (P < 0.05) higher milk SCC, neutrophil percent and significantly (P < 0.05) lower viability of both blood and milk neutrophils were observed in CM group of cows as compared to SCM and control group of cows. Phagocytic activity (PA) was significantly (P < 0.05) higher in milk neutrophils of SCM and CM cows as compared to control cows. Toll like receptors 2 and 4 in blood and milk neutrophils were found to be significantly (P < 0.05) higher, whereas, apoptosis in terms of altered mitochondrial transmembrane potential, Caspase 3 and 7 activities were found to be significantly (P < 0.05) lower in CM cows. Alterations in surface architecture of neutrophils in terms of formation of pseudopods was observed by scanning electron microscope (SEM) and found to be higher in CM cows. Blood neutrophils were found to be spherical as compared to milk neutrophils. Formation of neutrophil extracellular trap (NETs) were found milk neutrophils of CM cows, whereas, SCM and healthy cows did not exhibit NET formation. The study indicated a positive correlation between lower neutrophil apoptosis and higher expression of TLR2 and TLR4 with the formation of NETs and change in surface architecture. Formation of NET like structures seemed to be an effective mode of defense employed by neutrophils of cows suffering from clinical mastitis.