Anatomy and Physiology of Water Buffalo Mammary Glands: An Anatomofunctional Comparison with Dairy Cattle.

The present review aims to analyze the anatomical and physiological characteristics of the mammary gland and udders of water buffalo by making an anatomofunctional comparison with dairy cattle. It will also discuss the knowledge generated around the physiological regulation of milk ejection in the water buffalo. It was found that buffalo's average udder depth and width is approximately 20 cm smaller than Bos cattle. One of the main differences with dairy cattle is a longer teat canal length (around 8.25-11.56 cm), which highly influences buffalo milking. In this sense, a narrower teat canal (2.71 ± 0.10 cm) and thicker sphincter muscle are associated with needing higher vacuum levels when using machine milking in buffalo. Moreover, the predominant alveolar fraction of water buffalo storing 90-95% of the entire milk production is another element that can be related to the lower milk yields in buffalo (when compared to Bos cattle) and the requirements for prolonged prestimulation in this species. Considering the anatomical characteristics of water buffalo's udder could help improve bubaline dairy systems.

Viral Diseases in Water Buffalo (Bubalus bubalis): New Insights and Perspectives.

The water buffalo (Bubalus bubalis) has great adaptability to rustic environments and more variable conditions than cattle, who generally share the habitat. Diseases carried by buffaloes are relatively unknown and ignored and could be transmissible; an imbalance occurs between pathogens, environment, and susceptible hosts, generating a severe animal health problem. Also relevant is the effect of climate change on the populations of vectors that transmit viral diseases. The discovery of new virus variants that can pass from bovine (Bos) to buffalo or vice versa or to humans has highlighted the relevance of viruses crossing the host barrier. This review discusses the clinical viral diseases most reported in the water buffalo, characteristics, epidemiology, and recent findings about disease behavior, interaction with other species, the host, vectors, and pathogens. Diseases reviewed include Foot and Mouth Disease, Rinderpest, Malignant Catarrhal Fever, Infectious Bovine Rhinotracheitis, Bovine Viral Diarrhea, and Rabies. Also, vector-borne diseases include Lumpy Skin Disease, Ephemeral Fever, and Blue Tongue. The review also considers emerging viruses such as Buffalo Pox and Schmallenberg and, finally, other viruses such as papillomatosis. The knowledge and epidemiology of buffalo viral diseases must be constantly reconsidered and updated for adequate prevention and control programs.

Bacterial Meningitis in Buffaloes in Brazil.

Meningitis is the inflammation of the membranes surrounding the central nervous system and is poorly described in water buffaloes. Five cases of meningitis in adults buffaloes of the Murrah and Mediterranean breads were studied. All buffaloes came from a farm located in the municipality of Castanhal, Pará, Brazil at different times. Clinical examination showed neurological clinical signs, such as apathy, reluctance to move, spastic paresis especially of the pelvic limbs, hypermetria, difficulty getting up, pressing of the head into obstacles and convulsion. In three buffaloes, a large part of the horn had been lost, exposing the corresponding frontal sinus, through which a bloody to purulent exudate flowed. The hemogram revealed neutrophilic leukocytosis. At necropsy, adherence of the dura mater to the periosteum and a purulent to fibrinopurulent exudate were observed in the sulci of the cerebral cortex and on the pia mater over almost the entire surface of the brain and throughout the spinal cord. The cerebrospinal fluid had a cloudy aspect with fibrin filaments. The histopathology of buffaloes confirmed the diagnosis of bacterial fibrinopurulent meningitis. Buffaloes are susceptible to bacterial inflammation of the meninges due to fractures of the base of the horn and mostly present with neurological manifestations.

Functionally conserved PPARG exonic circRNAs enhance intramuscular fat deposition by regulating PPARG and HSL.

Circular RNAs (circRNA) are a kind of endogenous biological macromolecules that play significant roles in many biological processes, including adipogenesis, a precisely orchestrated process that is mediated by a large number of factors. Among them, peroxisome proliferator-activated receptor gamma (PPARG), is undoubtedly the most important regulator of adipocyte development in all types of adipose tissue. The formation of intramuscular fat (IMF), is a key factor that influences the meat quality in livestock animals. PPARG has been demonstrated to show a positive correlation with IMF deposition although the regulatory mechanism involved is not known. This study demonstrates that PPARG mediates IMF deposition by producing multiple exonic circRNAs (circPPARGs). Three circPPARGs promote adipogenic differentiation and inhibit the proliferation of intramuscular preadipocytes and these effects are conserved across several species including buffaloes, cattle and mice. Notably, circPPARG1 interacts with PPARG protein to inhibit the transcription of hormone sensitive lipase (HSL) involved in lipolysis. In addition, the positive effects of circPPARG1 on IMF deposition were identified in mice in vivo. Thus, PPARG drives IMF deposition, not only through the common transcription factor pathway, but also by producing circRNAs. This study provides new insights into our understanding of the regulatory mechanisms of PPARG in IMF deposition.

Trade and consumption of buffalo meat in Brazil.

Due to the lack of information concerning the buffalo production chain, the aim of this survey was to identify consumer behaviour, acceptance and trends in relation to buffalo meat. The survey research method was used, for which an online questionnaire was prepared using Google Forms, and distributed via digital platforms to Brazilian consumers. Using the exploratory technique of multivariate statistics, ordination was carried out by principal component analysis (PCA), where the words were grouped into dimensions. Of the respondents, 44.82% had already consumed buffalo meat; however, many did not know whether they had already tried it, since 50.24% of the respondents said they could not identify any difference between buffalo meat and beef. An identification and indication of the origin were considered important by 44.82% of respondents. The respondents cited 1130 different terms, the word 'quality' appearing in most responses. Principal component analysis explained 93.32% of the variations in the consumer perception of buffalo meat. A part of Brazilian consumers is unaware of the buffalo production chain, and have never had the opportunity to consume the meat. The creation and divulgation of a certificate of identification, together with the benefits to health from consuming buffalo meat is fundamental for consumer confidence, which, together with the ready availability of the product, would allow expansion, consolidation and improvement of the production chain. The results of the study show the potential for increasing the consumption of buffalo meat among Brazilian consumers, providing the product included a guarantee of origin, quality and healthiness.

Gestational monitoring in buffalo females: detection of Neospora caninum by PCR.

This study establishes the serological frequency against Neospora caninum on day zero and the presence of N. caninum DNA surveyed throughout the gestation of Bubalus bubalis females in a bovine buffalo system in the central zone of the state of Veracruz, Mexico. Blood samples were taken from 11 females in 6 different sampling periods and analyzed for N. caninum antibodies detection on day zero. DNA detection by PCR was performed on all sampling periods. The gestation months of the females were recorded for five trimesters by ultrasonography, as well as births and pregnancy losses. Recorded seropositivity and positivity for agent DNA were 90.9% (95% CI 58.7-9.7) and 36.3% (95% CI 10.9-69.2), respectively, on day zero. N. caninum DNA was detected between 18.1% (95% CI 2.3-51.7) and 45.4% (95% CI 16.7-76.6) over the five trimesters of observation, with three births and three abortions recorded. The studied water buffalo population had a high presence of N. caninum antibodies; however, the detection of N. caninum DNA remained below 47% in the females. The association was only observed in the detection of DNA with pregnant females (P 0.007). Our results support the hypothesis of the resistance of water buffaloes to infection and the onset of clinical signs against infection by N. caninum even upon a high possibility of infection and reinfection described in this production system in Mexico.

Biodiversity of Indonesian indigenous buffalo: First review of the status, challenges, and development opportunities.

In Indonesia, the buffalo is important for small and marginal farmers' livelihood and economic development as a source of food, working animal, and tourist attraction. Therefore, an in-depth study is needed to examine challenges and opportunities for buffalo development in Indonesia. In Indonesia, the buffalo is divided into two types: swamp buffalo and river buffalo. The buffalo population in Indonesia has declined significantly. A decrease of approximately 39.35% was recorded from 2022 to 2017. The decline occurred due to low reproduction rate and suboptimal rearing management systems. There are three buffalo-rearing systems: Intensive, semi-intensive, and extensive. The productivity of buffalo is diverse and closely related to the characteristics of the regional agroecosystem, consistent with existing natural resources and rearing management systems. The diversity of buffalo productivity provides a good opportunity to improve productivity. Improvement of buffalo genetics is urgently needed, by improving mating management, etc., especially to reduce potential inbreeding. In recent years, genetic and molecular research on Indonesian buffalo has made progress, including use of molecular markers, such as microsatellites and single-nucleotide polymorphisms, to evaluate genetic diversity within and among buffalo populations across Indonesia. In addition, studies are being conducted on the relationship of genotype mutations that contribute to appearance and phenotypic performance (heat stress, reproduction, behavior, coat color, and production attributes) in buffaloes. Identification of genetic diversity in local buffaloes can be improved using various genetic and genomic techniques. These findings will form a basis for the targeted conservation of local buffaloes in Indonesia. This study aimed to collect information on the genetic resources of the local buffalo, particularly its status and production system and provide recommendations for developing buffalo production in Indonesia.

Slaughtering of Water Buffalo (Bubalus bubalis) with and without Stunning: A Focus on the Neurobiology of Pain, Hyperalgesia, and Sensitization.

The slaughter process in livestock is considered a stressor where the transport and handling of animals, as well as the selected stunning and bleeding methods, can cause acute pain, distress, and suffering. In water buffaloes, although stunning is known to be performed before bleeding to induce unconsciousness, no emphasis is made on the nociceptive events during this process. Particularly, current mechanical stunning methods applied to cattle are unsuitable for water buffaloes due to anatomical differences in the skull from other large ruminants. Furthermore, although very high-pressure pneumatic (200-220 psi) may be effective in the frontal position for lighter-weight water buffalos, for heavier animals, it is less likely to be effective. The present review aims: (1) to analyze the anatomical particularities of water buffaloes to discuss the importance of selecting a stunning method suitable for buffaloes, and (2) to revise the potential pain-related consequences, such as hyperalgesia and sensitization, and the signs to assess the stun quality and death to comprehend the relevance of a proper technique according to the species.

A Novel Insight into the Identification of Potential SNP Markers for the Genomic Characterization of Buffalo Breeds in Pakistan.

Domestic buffaloes (Bubalus bubalis), known as water buffaloes, play a key role as versatile multipurpose agricultural animals in the Asiatic region. Pakistan, with the second-largest buffalo population in the world, holds a rich domestication history of buffaloes. The overall trends in buffalo production demand the genomic characterization of Pakistani buffalo breeds. To this end, the resequencing data of Pakistani breeds, along with buffalo breeds from 13 other countries, were retrieved from our previous study. This dataset, which contained 34,671,886 single-nucleotide polymorphisms (SNPs), was analyzed through a pipeline that was developed to compare possible allele differences among breeds at each SNP position. In contrast, other available tools only check for positional SNP differences for breed-specific markers. In total, 1918, 1549, 404, and 341 breed-specific markers were identified to characterize the Nili, Nili-Ravi, Azakheli, and Kundi breeds of Pakistani buffalo, respectively. Sufficient evidence in the form of phenotypic data, principal component analysis, admixture analysis, and linkage analysis showed that the Nili breed has maintained its distinct breed status despite sharing a close evolutionary relationship with the Nili-Ravi breed of buffalo. In this era of genome science, the conservation of these breeds and the further validation of the given selection markers in larger populations is a pressing need.

Strategies and Mechanisms of Thermal Compensation in Newborn Water Buffaloes.

Hypothermia is one of the principal causes of perinatal mortality in water buffaloes and can range from 3% to 17.9%. In ruminants, factors affecting hypothermia in newborns may be of intrinsic (e.g., level of neurodevelopment, birth weight, vitality score, amount of brown fat, skin features) or extrinsic origin (e.g., maternal care, environmental conditions, colostrum consumption). When newborn buffaloes are exposed to cold stress, thermoregulatory mechanisms such as peripheral vasoconstriction and shivering and non-shivering thermogenesis are activated to prevent hypothermia. Due to the properties of infrared thermography (IRT), as a technique that detects vasomotor changes triggered by a reduction in body temperature, evaluating the central and peripheral regions in newborn buffaloes is possible. This review aims to analyze behavioral, physiological, and morphological strategies and colostrum consumption as thermal compensation mechanisms in newborn water buffalo to cope with environmental changes affecting thermoneutrality. In addition, the importance of monitoring by IRT to identify hypothermia states will be highlighted. Going deeper into these topics related to the water buffalo is essential because, in recent years, this species has become more popular and is being bred in more geographic areas.

Predicting live weight using body volume formula in lactating water buffalo.

Live weight (LW) is an important piece of information within production systems, as it is related to several other economic characteristics. However, in the main buffalo-producing regions in the world, it is not common to periodically weigh the animals. We develop and evaluate linear, quadratic, and allometric mathematical models to predict LW using the body volume (BV) formula in lactating water buffalo (Bubalus bubalis) reared in southeastern Mexico. The LW (391.5 ± 138.9 kg) and BV (333.62 ± 58.51 dm3) were measured in 165 lactating Murrah buffalo aged between 3 and 10 years. The goodness-of-fit of the models was evaluated using the Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), coefficient of determination (R2), mean-squared error (MSE) and root MSE (RMSE). In addition, the developed models were evaluated through cross-validation (k-folds). The ability of the fitted models to predict the observed values was evaluated based on the RMSEP, R2, and mean absolute error (MAE). LW and BV were significantly positively and strongly correlated (r = 0.81; P < 0.001). The quadratic model had the lowest values of MSE (2788.12) and RMSE (52.80). On the other hand, the allometric model showed the lowest values of BIC (1319.24) and AIC (1313.07). The Quadratic and allometric models had lower values of MSEP and MAE. We recommend the quadratic and allometric models to predict the LW of lactating Murrah buffalo using BV as a predictor.

Assessment of thermal changes in water buffalo mobilized from the paddock and transported by short journeys.

Evaluating the welfare of buffaloes during transport is key to obtaining and commercializing high-quality meat products; however, effective assessments require recognizing several stressors that activate physiological mechanisms that can have repercussions on the health and productive performance of species. The aim of this study was to evaluate the surface temperatures of different body and head regions in this species during events prior, and posterior, to transport for short periods; that is, from paddock to loading. The second goal was to determine the level of correlation between thermal windows. This study used infrared thermography (IRT) to evaluate the surface temperature of 624 water buffaloes (Buffalypso breed) during 12 short trips (average duration = 2 h ± 20 min) by focusing on 11 regions of the body (Regio corporis), in the head regions (Regiones capitis) the face regions (Regiones faciei), Orbital region (Regio orbitalis) with special attention to structures such as the lacrimal caruncle, periocular area and lower eyelid (Regio palpebralis inferior); nasal region (Regio nasalis) with special attention to nostril thermal window; and regions of the skull (Regiones cranii) such as auricular region (Regio auricularis) with special attention to auditory canal and frontal-parietal region (Regio frontalis-parietalis) and trunk region (Truncus regionis) such as thoracic and abdominal regions, regions of the vertebral column (Columna vertebralis) with the thoracic vertebral region (Regio vertebralis thoracis) and lumbar region (Regio lumbalis); and regions of the pelvis limb (Regiones membri pelvini). Recordings were made during seven phases: paddock (P1), herding (P2), corral (P3), chute handling (P4), shipping (P5), pre- (P6), and post-transport (P7). A total of 48,048 readings were obtained from 11 thermal windows. The results showed that the surface temperatures of the windows increased by as much as 5°C during P2, P3, P5, P6, and P7 compared to P1 and P4 (p < 0.0001). Differences of at least 1°C were also observed between thermal windows in the craniofacial, lateral corporal, and peripheral zones (p < 0.0001). Finally, a strong positive correlation (r = 0.9, p < 0.0001) was found between the thermal windows. These findings lead to the conclusion that the surface temperature of the craniofacial and corporal regions of buffaloes transported for short periods varied in relation to the phase of mobilization (from paddock to post-transport), likely as a response to stressful factors, since herding and loading increased the thermal values in each window. The second conclusion is that there are strong positive correlations between central and peripheral thermal windows.

Diversity in Cell Morphology, Composition, and Function among Adipose Depots in River Buffaloes.

Fat deposition is a significant economic trait in livestock animals. Adipose tissues (ATs) developed in subcutaneous and visceral depots are considered waste whereas those within muscle are highly valued. In river buffaloes, lipogenesis is highly active in subcutaneous (especially in the sternum subcutaneous) and visceral depots but not in muscle tissue. Revealing the features and functions of ATs in different depots is significant for the regulation of their development. Here, we characterize the cell size, composition, and function of six AT depots in river buffaloes. Our data support that the subcutaneous AT depots have a larger cell size than visceral AT depots, and the subcutaneous AT depots, especially the sternum subcutaneous AT, are mainly associated with the extracellular matrix whereas the visceral AT depots are mainly associated with immunity. We found that sternum subcutaneous AT is significantly different from ATs in other depots, due to the high unsaturated fatty acid content and the significant association with metabolic protection. The perirenal AT is more active in FA oxidation for energy supply. In addition, the expression of HOX paralogs supports the variable origins of ATs in different depots, indicating that the development of ATs in different depots is mediated by their progenitor cells. The present study enhances our understanding of the cellular and molecular features, metabolism, and origin of AT depots in buffaloes, which is significant for the regulation of fat deposition and provides new insights into the features of AT depots in multiple discrete locations.

Assessment of antioxidant enzymes during the development of the digenetic trematode parasite Gastrothylax crumenifer, infecting the rumen of Indian water buffalo, Bubalus bubalis.

The presence of antioxidant enzymes in helminth parasites is well known. These enzymes help the parasites to survive in their hosts by detoxifying host-generated reactive oxygen species (ROS). The literature survey reveals that most of the studies related to antioxidant enzymes in helminth parasites are restricted to the adult stage while the larval stages are neglected. The present investigation is designed to evaluate the level of antioxidant enzymes in the adult and larval stages of rumen-infecting paramphistome parasites, Gastrothylax crumenifer. The larval stages include 0-day eggs, 4-day eggs, and eggs containing mature miracidia, cercariae, and metacercariae. Antioxidant enzyme assays were performed using standard assay protocols. Our findings revealed an increasing pattern in the level of Glutathione-S-Transferase (GST), Superoxide Dismutase (SOD), Glutathione Reductase (GR), and Glutathione Peroxidase (GPx) antioxidant enzymes during the development from 0-day eggs to the adult stage. Overall analysis shows that adult worms have higher antioxidant enzyme activity as compared to the larval stages, indicating that adult flukes are more adapted to oxidative stress. It can be concluded that the miracidia, cercarial, and metacercarial developmental stages of G. crumenifer possess a considerable level of antioxidant enzymes suitable to overcome the oxidative stress encountered during the development and help them in the completion of the life cycle and survival in the definitive host.

Estimation of body weight using body measurements in female water buffaloes reared in southeastern Mexico.

Buffalo farming is an important livestock activity in Mexico. However, the low technological level of the farms makes it difficult to monitor the growth rates of the animals. The objectives of this study were to analyse the body measurements of 107 adult female Murrah buffaloes, to estimate the interrelationships between those measurements and body weight, and to develop equations to predict body weight (BW) using body measurements including withers at height (WH), rump height (RH), body height (BH), heart girth (HG), abdominal girth (AG), pelvic girth (PG), body length (BL), girth circumference (GC), diagonal body length (DBL), pelvic circumference (PC), and abdomen circumference (AC). The study was conducted on two commercial farms in southern Mexico. Pearson correlation and stepwise regression techniques were used for the data analysis. To find out the best regression models, we used model quality criteria such as coefficient of determination (R2), adjusted R2 (Adj.R2), root mean square error (RMSE), Mallow's Cp, Akaike's information criteria (AIC), Bayesian information criteria (BIC), and coefficient of variation (CV). Correlation results indicated that BW had a positive high correlation (P < 0.01) of all the measured traits. Model 4 (-780.56 + 311.76GC + 383.51DBL + 51.82PC + 47.65AC-106.78BL) was the best regression model with a higher R2 (0.87), Adj. R2 (0.86) smaller Cp (4.24), AIC (749.19), BIC (752.16), and RMSE (36.91). The current study suggests that GC, DBL, PC, AC, and BL might be used in combination to estimate BW of adult female Murrah buffaloes.

Flow cytometric detection of IFN-γ production and Caspase-3 activation in CD4+ T lymphocytes to discriminate between healthy and Mycobacterium bovis naturally infected water buffaloes.

Tuberculosis has a negative economic impact on buffalo farming, and it poses a potential threat to human health. Interferon-gamma (IFN-γ) plays a central role in protection against mycobacterial diseases, illustrating the importance of T-cell mediated immune responses in tuberculosis infection. Recently, the expression of Caspase-3, a critical executor of apoptosis, in M. tuberculosis-specific IFN-γ+CD4+ T cells was used as a new marker to distinguish active from latent tuberculosis infection in humans. The aims of this work were to develop a whole blood flow cytometric assay to detect the production of IFN-γ and the activation of Caspase-3 by CD4+ T lymphocytes from water buffalo and to evaluate whether these parameters can discriminate between healthy and M. bovis naturally infected buffaloes. A total of 35 Italian Mediterranean buffaloes were grouped in two groups: uninfected and M. bovis infected (based on the results of antemortem diagnostic tests: single intradermal tuberculin (SIT) and ELISA IFN-γ tests). Whole blood was incubated for 6 h with tubercular antigens: PPD-B, PPD-A, ESAT-6/CFP-10 and a new mix of precocious secreted antigens (PA). Our results showed a significant increase in the percentage of IFN-γ+CD4+ T cells in infected compared to the uninfected animals after each stimulus. Improved sensitivity of the assay was obtained by including the stimulation with the new mix of PA. Interestingly, we observed a concomitant decrease in percentage of Caspase-3+CD4+ T cells in M. bovis infected animals compared to the control healthy ones, regardless of the stimulus used. Overall, these results showed that M. bovis infection activates CD4+ T lymphocytes to produce IFN-γ and at the same time causes a concomitant decrease of Caspase-3 activation in CD4+ T cells. This study for the first time in water buffalo describes the development of a whole blood flow cytometric assay for the detection of IFN-γ producing CD4+ T cells and proposes the expression of active Caspase-3 as an additional bovine TB biomarker. Although further studies are needed to better understand the mechanisms of Caspase-3-mediated cell death during tuberculosis, our data can help to better understand the cellular immune response to M. bovis infection in buffalo species.

Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood.

Monocytes are innate immune system key players with pivotal roles during infection and inflammation. They migrate into tissues and differentiate into myeloid effect cells (macrophages, dendritic cells) which orchestrate inflammatory processes and are interfaces between the innate and adaptive immune responses. Their clinical relevance to health and disease of cattle (Bos taurus) and water buffalo (Bubalus bubalis), two of the most important livestock species, has been highlighted in physiologic (pregnancy) and pathologic (mastitis, metritis, and viral infections) conditions. The existence of three different monocyte subsets in cattle was established by flow cytometry (FC), as follows: classical (cM; CD14++ CD16-/low ), intermediate (intM; CD14++/+ CD16+ ), and non-classical (ncM; CD14-/low CD16++ ) monocytes. FC applications for studying the immune system of cattle and water buffalo still have significant limitations. In this article, we describe some practical approaches to overcome these limitations and, in particular, allow the identification and enumeration of cM, intM, and ncM subpopulations in cattle and buffalo peripheral blood. Indeed, we propose the new procedure lyse/wash/no-centrifugation (L/W/NC) that can be combined with the FC absolute counting procedures and can overcome specific issues of the lyse/no-wash protocols (L/NW). Finally, for the first time, we demonstrated the existence of cM, intM, and ncM monocyte subsets also in the water buffalo, showing some interesting differences with cattle, such as the bubaline cM are mainly CD14+/++ /CD16+ . These subtle differences may influence inflammatory disease regulation in, for example, mastitis and metritis. The upregulation of CD16 expression on cM may reveal different monocyte priming, leading to different functional features of macrophages/dendritic cells in tissues after infection. © 2023 Wiley Periodicals LLC. Basic Protocol: Absolute count of cM, intM, and ncM without compensation Alternate Protocol: Absolute count of cM, intM, and ncM for single laser platform Support Protocol 1: In-house monoclonal antibody labeling using a Pacific Blue™ kit Support Protocol 2: In-house monoclonal antibody labeling using an Alexa Fluor® 647 kit Support Protocol 3: Titration of fluorochrome-conjugated antibodies.

In-depth immunophenotyping reveals significant alteration of lymphocytes in buffalo with brucellosis.

Water buffalo (Bubalus bubalis) has a prominent position in the livestock industry worldwide but still suffers from limited knowledge on the mechanisms regulating the immune against infections, including brucellosis (BRC), one of the most significant neglected zoonotic diseases of livestock. Seventy-three buffalo were recruited for the study. Thirty-five were naturally infected with Brucella spp. The aims of the study were to (i) verify the cross-reactivity of 16 monoclonal antibodies (mAbs) developed against human, bovine, and ovine antigens; (ii) evaluate lymphocyte subset alterations in BRC positive buffalo; (iii) evaluate the use of the canonical discriminant analysis (CDA), with flow cytometric data, to discriminate BRC positive from negative animals. A new set of eight mAbs (anti CD3e, CD16, CD18, CD45R0, CD79a; CD172a) were shown to cross-react with water buffalo orthologous molecules. BRC positive animals presented a significant (p < 0.0001) decrease in the percentage of PBMC (29.5 vs. 40.3), total, T and B lymphocytes (23.0 vs. 35.5, 19.2 vs. 28.9, 2.6 vs. 5.7, respectively). In contrast, they showed an increase in percentage of granulocytes (65.2 vs. 55.1; p < 0.0001) and B lymphocytes CD21neg (22.9 vs. 16.1; p = 0.0067), a higher T/B lymphocyte ratio (10.3 vs. 6.4; p = 0.0011) and CD3+ /CD21+ (14.7 vs. 8.3; p = 0.0005) ratio. The CDA, applied to 33 different flow cytometric traits, allowed the discrimination of all BRC positive from negative buffalo. Although this is a preliminary study, our results show that flow cytometry can be used in a wide range of applications in livestock diseases, including in support of uncertain BRC diagnoses.

A long non-coding RNA lnc210 promotes adipogenic differentiation of buffalo intramuscular adipocytes.

Intramuscular fat (IMF) content is one of the most significant factors influencing beef quality in terms of tenderness, flavor, and juiciness. Thus, internal factors affecting IMF deposition have received considerable attention for decades. In this study, we demonstrated a long non-coding RNA, lnc210, promoted adipogenic differentiation of buffalo intramuscular adipocytes. lnc210 was rich in adipose tissue and showed increased expression with the adipogenic differentiation of buffalo intramuscular adipocytes. lnc210 was mainly expressed in the nucleus of adipocytes. Full-length lnc210 was obtained by rapid amplification of cDNA ends technology. lnc210 overexpression promoted lipid accumulation by upregulating the mRNA expression of peroxisome proliferator-activated receptor gamma (PPARG) and CCAAT enhancer binding protein alpha (C/EBPα) in buffalo intramuscular adipocytes. These results provide a basis for an in-depth analysis of the role of lnc210 in accelerating IMF deposition in buffaloes.

Random regression models using B-splines functions provide more accurate genomic breeding values for milk yield and lactation persistence in Murrah buffaloes.

There is a great worldwide demand for cheese made with buffalo milk, due to its flavour and nutritional properties. In this context, there is a need for increasing the efficiency of buffalo milk production (including lactation persistence), which can be achieved through genomic selection. The most used methods for the genetic evaluation of longitudinal data, such as milk-related traits, are based on random regression models (RRM). The choice of the best covariance functions and polynomial order for modelling the random effects is an important step to properly fit RRM. To our best knowledge, there are no studies evaluating the impact of the order and covariance function (Legendre polynomials-LEG and B-splines-BSP) used to fit RRM for genomic prediction of breeding values in dairy buffaloes. Therefore, the main objectives of this study were to estimate variance components and evaluate the performance of LEG and BSP functions of different orders on the predictive ability of genomic breeding values for the first three lactations of milk yield (MY1, MY2, and MY3) and lactation persistence (LP1, LP2, and LP3) of Brazilian Murrah. Twenty-two models for each lactation were contrasted based on goodness of fit, genetic parameter estimates, and predictive ability. Overall, the models of higher orders of LEG or BSP had a better performance based on the deviance information criterion (DIC). The daily heritability estimates ranged from 0.01 to 0.30 for MY1, 0.08 to 0.42 for MY2, and from 0.05 to 0.47 for MY3. For lactation persistence (LP), the heritability estimates ranged from 0.09 to 0.32 for LP1, from 0.15 to 0.33 for LP2, and from 0.06 to 0.32 for LP3. In general, the curves plotted for variance components and heritability estimates based on BSP models presented lower oscillation along the lactation trajectory. Similar predictive ability was observed among the models. Considering a balance between the complexity of the model, goodness of fit, and credibility of the results, RRM using quadratic B-splines functions based on four or five segments to model the systematic, additive genetic, and permanent environment curves provide better fit with no significant differences between genetic variances estimates, heritabilities, and predictive ability for the genomic evaluation of dairy buffaloes.