Water Buffalo Bubalus Bubalis Research Articles

Transcriptional Dynamics of Homeobox C11 Gene in Water Buffalo Bubalus bubalis

The Hox complex contains 39 genes clustered into four groups involved in cell differentiation and development. We cloned full-length sequence of Hoxc11 gene from water buffalo Bubalus bubalis, assessed its copy number, localized the same onto the chromosome 5, and studied its evolutionary conservation across the species. Northern hybridization of Hoxc11 showed a 2.2 kb band in the tissues analyzed. Real-Time PCR showed highest expression of Hoxc11 gene in lung followed by spleen, spermatozoa, and testis. Six interacting partners of this gene showed higher expression in spleen, lung, testis, and spermatozoa. During the early stages of development, Hoxc11 and its interacting partners both showed lower expression, which then became prominent during the age of 1-3 years, regressed drastically thereafter, and remained so until the animal's life time (∼ 20 years). The high expression of Hoxc11 and its interacting partners in spermatozoa and testis during the onset of puberty suggests its likely role in the differentiation of gonads and subsequent reproductive activities. Additional work on Hoxc11 especially, in the context of respiratory, immunological, and in/fertility in other species, including humans would be useful for establishing its broader biological significance towards the enrichment of functional and comparative genomics.

A diagnostic protocol to identify water buffaloes (Bubalus bubalis) vaccinated with Brucella abortus strain RB51 vaccine.

The use of live vaccine strain RB51 for vaccination of domestic water buffaloes (Bubalus bubalis) at risk of infection with Brucella abortus is permitted notwithstanding the plans for the eradication and only under strict veterinary control. The antibodies induced by RB51 vaccination are not detectable using conventional diagnostic techniques; therefore, it is necessary to have a specific diagnostic tool able to discriminate vaccinated from unvaccinated animals. The combination of a complement fixation test (CFT) with specific RB51 antigen (RB51-CFT) and a brucellin skin test has been demonstrated to be a reliable diagnostic system to identify single cattle (Bos taurus) vaccinated with RB51. So far, no data are available in the international scientific literature regarding the use of this test association in water buffalo. For this reason the suitability of this test combination has been evaluated in a water buffalo herd. One hundred twenty-seven animals farmed in a herd of Salerno province (Campania, Southern Italy), in the context of a presumptive unauthorized use of RB51 vaccine were chosen for this study. All tested animals resulted negative to Rose Bengal test (RBT) and complement fixation test (CFT) used for the detection of specific antibodies against Brucella field strains. Seventy-one animals (56%) developed RB51 antigen-specific CFT (RB51-CFT) antibodies against RB51 vaccine in a first sampling, while 104 animals (82%) gave positive result to a second serum sampling conducted 11 days after the intradermal inoculation of the RB51 brucellin. One hundred and seven animals (84%) showed a positive reaction to the RB51-CFT in at least 1 sampling, while 111 animals (87%) resulted positive to the RB51 brucellin skin test. Thus, analysing the results of the 3 testing in parallel, 119 animals (94%) were positive to at least 1 of the performed tests. The results suggest that the use in parallel of the RB51 brucellin skin test with RB51-CFT may represent a reliable diagnostic system to identify water buffaloes vaccinated with RB51 vaccine.

Differentiated evolutionary relationships among chordates from comparative alignments of multiple sequences of MyoD and MyoG myogenic regulatory factors.

MyoD and MyoG are transcription factors that have essential roles in myogenic lineage determination and muscle differentiation. The purpose of this study was to compare multiple amino acid sequences of myogenic regulatory proteins to infer evolutionary relationships among chordates. Protein sequences from Mus musculus (P10085 and P12979), human Homo sapiens (P15172 and P15173), bovine Bos taurus (Q7YS82 and Q7YS81), wild pig Sus scrofa (P49811 and P49812), quail Coturnix coturnix (P21572 and P34060), chicken Gallus gallus (P16075 and P17920), rat Rattus norvegicus (Q02346 and P20428), domestic water buffalo Bubalus bubalis (D2SP11 and A7L034), and sheep Ovis aries (Q90477 and D3YKV7) were searched from a non-redundant protein sequence database UniProtKB/Swiss-Prot, and subsequently analyzed using the Mega6.0 software. MyoD evolutionary analyses revealed the presence of three main clusters with all mammals branched in one cluster, members of the order Rodentia (mouse and rat) in a second branch linked to the first, and birds of the order Galliformes (chicken and quail) remaining isolated in a third. MyoG evolutionary analyses aligned sequences in two main clusters, all mammalian specimens grouped in different sub-branches, and birds clustered in a second branch. These analyses suggest that the evolution of MyoD and MyoG was driven by different pathways.

Purification and characterization of xanthine oxidase from liver of the water buffalo Bubalus bubalis

Xanthine oxidase (XO) is an important enzyme with broad medical applications as detecting reagents in many diagnostic kits. In this study, buffalo liver xanthine oxidase (BLXO) was purified to homogeneity by acetone precipitation and chromatography on DEAE-cellulose and Sephacryl S-300 columns with a specific activity of 7.2 units / mg protein which represent 31.3 folds. The native molecular weight of the purified enzyme is 200 kDa and its subunit molecular weight was determined by SDS-PAGE to be 67 kDa. The isoelectric point (pI) value of BLXO isoenzyme is at pH 6.0 – 6.2. It displayed its pH optima at 7.6 and the Km value is 1.1 mM xanthine. FeCl2 increased the activity of BLXO while CuCl2, MnCl2 and ZnCl2 were found to be inhibitors of the purified enzyme. Allopurinol inhibits BLXO competitively and has one binding site on it with Ki value of 0.025mM.

2D-PAGE analysis of the soluble proteins of the tropical liver fluke, Fasciola gigantica and biliary amphistome, Gigantocotyle explanatum, concurrently infecting Bubalus bubalis.

The digenetic trematodes, Fasciola gigantica and Gigantocotyle explanatum, belonging to the family Fasciolidae and Paramphistomidae respectively, have been often found to concurrently infect the liver of Indian water buffalo Bubalus bubalis, causing serious pathological damage to the vital organ, incurring huge economic losses. In the present study the soluble gene products of both F. gigantica and G. explanatum were analyzed by 2 dimensional polyacrylamide gel electrophoresis. The soluble proteomic profile revealed considerable similarity as well as differences in the size, distribution pattern, total number, the isoelectric point (pI) and molecular weight (Mr) of the resolved polypeptide spots. The maximum number of polypeptide spots with a molecular weight range of >10 to 160kDa were recorded with a pI range of 7-9 followed by pI range of 5-7, 9-10 and 3-5 in both the parasites. However, considerable variation was recorded in the Mr of the polypeptides belonging to each pI range. The genetic heterogeneity could be an obvious contributing factor for such differences but some polypeptides appeared to be conserved in the two species. The molecular similarities and the habitat preference by these worms may be a consequence of microenvironmental cues that guide these flukes to reach their habitat through different routes and establish a successful host-parasite relationship.

Comparison of water buffalo (Bubalus bubalis) with crossbred and purebred Brahman cattle for growth performance on savannah and slaughter traits at four ages in Venezuela

Males (n = 132) of water buffalo (Buffalo), Brahman crossbred and purebred cattle (Brahman) were born, raised, weaned, castrated (half of the animals at 7 months) and fattened on the same grazing savannahs, and then slaughtered at four different ages (7, 17, 19 and 24 months) to compare growth and slaughter traits and examine post-weaning castration effects. Buffalo showed consistently faster weight gains and heavier carcasses but had lower dressing percentages than Brahman cattle due to higher proportions of hide, head, and red and white viscera at each age (P < 0.05). Additionally, Buffalo had abundant fat cover distribution, and whiter subcutaneous fat than Brahman (P < 0.05), this resulted in a higher proportion of Buffalo carcasses that graded in the second quality Venezuelan grade ‘A’ (P < 0.05). Regardless of species, bulls had higher average daily gain and liveweight than steers (P < 0.05); however, no differences were detected in carcass weight and dressing percentages (P > 0.05). Also, at the same weight steers produced carcasses with thicker back fat, more abundant fat cover distribution and whiter fat than bulls (P < 0.05). With respect to age groups, there were concomitant increases in slaughter weight, carcass weight, and dressing percentage as age increased (P < 0.01). As age advanced, carcasses had a more convex leg profile, abundant fat cover, thicker back fat, higher marbling and greater REA (P < 0.05). Based on the observed growth and carcass traits, Buffalo offer a promising alternative for meat production under savannah feeding production systems.

Molecular investigation of foot-and-mouth disease virus in domestic bovids from Gharbia, Egypt

An outbreak of foot-and-mouth disease (FMD) affecting cattle and water buffalo (Bubalus bubalis) occurred in Egypt during 2012/2013. The present study was undertaken to determine the current strains of the FMD virus (FMDV) and the prevalence of FMD among cattle and buffalo in Gharbia, Egypt. The diagnostic sensitivity of two RT-PCR assays for the detection of FMDV was evaluated. The results revealed that SAT2 was the causative agent. The percentage of infected of animals varied with the detection method, ranging from 62.5 % by the untranslated region (UTR) RT-PCR to 75.6 % by SAT2 RT-PCR. The overall prevalence and mortality rates were 100 and 21 %, respectively. The mortality was higher in buffalo (23.3 %) than it was in cattle (17 %). A partial sequence of SAT2 was identical (90-100 %) to Egyptian isolates and was close in similarity to sequences from Sudan and Libya. In conclusion, FMD in Egypt is caused by SAT2. No other serotypes were detected. The results of this study provided the valuable data regarding the epidemiology of SAT2 in cattle and water buffalo from Egypt, which strengthens the need to change the strategies of both control and prevention that help to prevent the spread of the disease.

Differential expression of Homeobox C11 protein in water buffalo Bubalus bubalis and its putative 3D structure.

BackgroundThe Homeobox (Hox) family complex contains 39 genes, clustered into four groups (A-D) all expressing in sequential manner. The HOX proteins are transcriptional factors involved in regulation of pattern formation of the anterio-posterior body axis across the species. Most of the Hox family genes have been studied with respect to their organization and expression during the embryonic stages. However, expression pattern of Homeobox C11 (Hoxc11) gene in the 5′ region, particularly in higher mammals remains largely unexplored.ResultsWe cloned and expressed Homeobox C11 (Hoxc11) gene from water buffalo Bubalus bubalis. The recombinant HOXC11 protein expressed as inclusion bodies was solubilized in Tris buffer (10 mM, pH-6.5) and purified using Ni-NTA affinity column. The purity and molecular weight of HOXC11 protein (~33 kDa) were confirmed by SDS-PAGE and western blot analysis. Employing immunohistochemistry approach, we localized HOXC11 protein in the nuclei across the tissues of buffalo. Western blot analysis showed highest expression of HOXC11 protein in kidney and lung although its possible renal and respiratory roles are not yet established. Electrophoretic mobility shift assay (EMSA) demonstrated the specific binding of HOXC11 protein with the promoter element, CE-LPH1 of lactase-phlorizin hydrolase (LPH) gene showing reduced mobility of the protein-DNA complex, corroborating with earlier report on the possible role of this protein in intestinal functions. In silico analysis of HOXC11 showed predominance of α helices and presence of six conserved domains. We deduced the putative 3D structure of HOXC11 protein and fifteen possible DNA interacting residues within the homeodomain.ConclusionsPresent study augments our understanding on the specific expression of HOXC11 protein in kidney and lung in water buffalo. The fifteen DNA interacting residues reported herein provide an opportunity to establish much broader structural and functional perspectives of HOXC11 protein in the context of genome analysis in general and animal biotechnology in particular.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-638) contains supplementary material, which is available to authorized users.

Forest without prey: livestock sustain a leopard Panthera pardus population in Pakistan

AbstractHuman–carnivore conflict is one of the major challenges in the management of populations of large carnivores. Concerns include the increasing human population; habitat loss as a result of degradation and fragmentation of forest; and livestock predation as a result of a lack of natural prey, leading to retaliatory killings of wild carnivores. Conflicts may be further aggravated by occasional attacks that result in injury and loss of human life. The level of consumption of prey species by a predator is a benchmark to evaluate the scale of this conflict. We used a newly developed DNA-based diet analysis to study the prey profile of common leopards Panthera pardus in Ayubia National Park, Pakistan. The results suggest that the common leopard is a generalist predator, subsisting mainly on domestic animals. Based on the frequency of occurrence of prey items in 57 faecal samples, the diet of the leopard is dominated by domestic goat Capra hircus (64.9%), followed by domestic dog Canis lupus familiaris (17.5%) and cow Bos taurus (12.3%). Domestic animals (goat, dog, cow, water buffalo Bubalus bubalis, horse Equus caballus and sheep Ovis aries) occurred in 54 (95%) of the 57 samples. We recommend a two-step strategy to mitigate this conflict: (1) introducing incentives for increased acceptance of leopards among local communities in the vicinity of the protected area and (2) increasing the availability of wild prey. We hope that the results of this study will contribute to the survival of the leopard in Pakistan.

Molecular characterisation of T cell receptor-zeta subunit (CD247) gene in buffalo (Bubalus bubalis)

The cDNA encoding T cell receptor-zeta (TCR-ζ; CD247) molecule of water buffalo (Bubalus bubalis) was isolated, cloned and sequenced in the present study. The CD247 cDNA comprised 1078 nucleotides including a 30 nucleotide 5′-untranslated region (UTR), 495 nucleotide single open reading frame (ORF) and 553 nucleotide 3′-UTR. Deduced amino acid of buffalo CD247 sequence was two residues shorter than the corresponding cattle and sheep sequences. However, ruminant-specific insertions and substitutions in intra-cytoplasmic (IC) domain were present in buffalo. Immunoreceptor tyrosine-based activation motifs (ITAMs), the important motifs for TCR signalling, were totally conserved among ruminants including buffalo. The 3′-UTR region of the buffalo CD247 was highly homologous to the corresponding region in the cattle sequence and showed lack of polymorphism after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using HaeIII and MseI restriction enzymes in buffalo population. Phylogenetically, buffalo sequence was closer to cattle sequence under the ruminant's lineage. The conserved nature of this gene ensures TCR integrity which is vital for induction of optimal and efficient immune response.

Propofol and thiopentone sodium as induction agents in water buffaloes (Bubalus bubalis): a comparative study

Propofol and thiopentone sodium as induction agents were evaluated in 12 healthy water buffaloes of either sex (3–5 years; 350–500 kg). Midazolam (0.2 mg/kg) + butorphanol (0.02 mg/kg) intravenously was used for premedication in both groups. After 5 min of premedication, anaesthesia was induced by 1% propofol (3 mg kg−1) in PRO group and by 5% thiopentone sodium (10 mg kg−1) in THIO group and maintained with isoflurane-oxygen mixture up to 30 min. Anaesthetic efficacy was assessed on the basis of quality of induction and recovery, time taken for induction and recovery and incidence of side effects. Heart rate, respiration rate, rectal temperature and mean blood pressure were also monitored. Propofol provided rapid and smooth induction of anaesthesia along with satisfactory conditions for endotracheal intubation. Propofol was associated with lesser respiratory depression than thiopentone sodium. Side effects like apnea and regurgitation were lesser and recovery was faster and smoother following propofol administration. Propofol was shown to be superior than thiopentone sodium and can be used as an alternative to thiopentone sodium for induction of general anaesthesia in water buffaloes.

Traumatic Reticuloperitonitis in Water Buffalo (Bubalus bubalis): Clinical Findings and the Associated Inflammatory Response

The present study was carried out to describe the clinical picture of traumatic reticuloperitonitis (TRP) in water buffalo (Bubalus bubalis) and to evaluate the inflammatory and immunologic responses for this clinical condition. Twenty-two buffalo with acute local TRP were monitored in our study. Additionally, 10 clinically healthy buffalo were randomly selected and served as controls. Acute local TRP was initially diagnosed by clinical examination and confirmed by ultrasonographic (USG) examination and/or necropsy findings. Blood samples were collected from all examined buffalo to measure the respective levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, IL-10 and interferon gamma (INF)-γ, serum amyloid A (SAA), C-reactive protein (CRP), haptoglobin (Hp), fibrinogen (Fb), and serum sialic acid (SSA). It was found that TNF-α, IL-1β, IL-6, IL-10, SAA, CRP, Hp, Fb, and SSA were significantly higher in buffalo with TRP than the controls. Our findings suggest that the examined immunologic variables were helpful in documenting the inflammatory response in buffalo with TRP. However, their diagnostic usefulness only becomes apparent when considered in tandem with the clinical findings for any given animal, its anamnesis, and a subsequent USG assessment. Due to the frequent complications of TRP, more accurate indicators of its occurrence and severity would be useful.

The efficacy and safety of alphacypermethrin as a pour-on treatment for water buffalo (Bubalus bubalis) infested with Haematopinus tuberculatus (Phthiraptera: Haematopinidae)

The sucking louse Haematopinus tuberculatus (Burmeister 1839) is an ectoparasite of buffaloes, cattle, camels, and American bison. Alphacypermethrin (ACYP) is a pyrethroid insecticide commonly used to control arthropods of veterinary and public health interest. Therapeutics, such as antiparasitic compounds, is often administered to buffaloes based on dosage and intervals recommended for cattle because very few drugs have buffalo-specific label indications. A trial was conducted on 20 louse-infested buffaloes at a farm to assess the efficacy and safety of ACYP pour-on, at the manufacturer's recommended dose for cattle, on buffaloes naturally infested by H. tuberculatus. Ten animals were assigned to ACYP-treated group (ACYP-group) and ten to untreated control group (C-group). On day 0, all ACYP-group buffaloes received alphacypermethrin pour-on. Louse counts were performed on days -1, 7, 14, 21, 28, 35, 42, 49, and 56 at eight predilection sites on the skin of each buffalo. ACYP was completely effective (100%) at day 7, highly effective (99.8%) at day 14, and completely effective (100%) from day 21 until the end of the study (day 56 post-treatment). During the trial, ACYP was well tolerated by all animals as there were no observed clinically adverse reactions. The results of this trial suggest that ACYP is an effective, safe, and user-friendly compound suitable for treatment of buffaloes with natural louse infestations.

Monitoring the ungulate prey of the Komodo dragonVaranus komodoensis: distance sampling or faecal counts?

Monitoring the abundances of prey is important for informing the management of threatened and endangered predators. We evaluated the usefulness of faecal counts and distance sampling for monitoring the abundances of rusa deerRusa timorensis, feral pigSus scrofaand water buffaloBubalus bubalis, the three key prey of the Komodo dragonVaranus komodoensis, at 11 sites on five islands in and around Komodo National Park, eastern Indonesia. We used species‐specific global detection functions and cluster sizes (i.e. multiple covariates distance sampling) to estimate densities of rusa deer and feral pig, but there were too few observations to estimate densities of water buffalo. Rusa deer densities varied from from 2.5 to 165.5 deer/km2with coefficients of variation (CVs) of 15‐105%. Feral pig densities varied from 0.0 to 25.2 pigs/km2with CVs of 25‐106%. There was a positive relationship between estimated faecal densities and estimated population densities for both rusa deer and feral pig: the form of the relationship was non‐linear for rusa deer, but there was similar support for linear and non‐linear relationships for feral pig. We found that faecal counts were more useful when ungulate densities were too low to estimate densities with distance sampling. Faecal count methods were also easier for field staff to conduct than distance sampling. Because spatial and temporal variation in ungulate density is likely to influence the population dynamics of the Komodo dragon, we recommend that annual monitoring of ungulates in and around Komodo National Park be undertaken using distance sampling and faecal counts. The relationships reported here will also be useful for managers establishing monitoring programmes for feral pig, rusa deer and water buffalo elsewhere in their native and exotic ranges.

Estimated Prevalence of Johne’s Disease in Herds of Water Buffaloes (Bubalus Bubalis) in the Province of Caserta

Paratuberculosis is a chronic infection of domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis. The persistence of paratuberculosis infection for months up to years without exhibiting any clinical signs makes the diagnosis and control program a difficult proposition. Limited informations on prevalence of paratuberculosis in water buffaloes (Bubalus bubalis) are available. We carried out a study on 1350 buffaloes belonging to 56 herds in the Caserta province, of Campania region, Italy. The prevalence of infected buffalo dairy herds was estimated by a commercial ELISA kit of individual blood samples of animals over 24 months of age. On the basis of performance (sensitivity 43%, specificity 99.3%) of ELISA test on serum, the resulting true prevalence at animal level and at herd level was 4% (95% CI 3% to 5%) and 74,1% (95% CI 71.8% to 76%). Considering the paucity of epidemiological reports in the region our results could be a useful contribution towards the prevention of buffalo paratuberculosis in the area.

Cellular Immune Response in Water Buffalo Placentas After Inoculation with Neospora caninum During Early Gestation

• Water buffalo (Bubalus bubalis) play a crucial role in tropical areas and their importance is increasing in western nations • Susceptibility to Neospora caninum (N. caninum) is not fully understood, although abortion has been confirmed in buffalos, and an exacerbated cellular immune response was associated with bovine abortion (1, 2, 3) • The aim of this work was to characterize the placental immune response following infection of pregnant Mediterranean water buffalo

Immunodetection of coproantigens for the diagnosis of amphistomosis in naturally infected Indian Water Buffalo, Bubalus bubalis

The infection of gastrointestinal helminths in livestock is routinely diagnosed by microscopical examination of faecal samples for the presence of ova/eggs but this approach becomes ineffective for the seasonally egg producing trematodes. Therefore, an alternative approach to detect the coproantigens of liver and rumen amphistomes, Gigantocotyle explanatum and Gastrothylax crumenifer respectively, infecting Indian water buffalo Bubalus bubalis, was undertaken using ELISA, immunodot and countercurrent immunoelectrophoresis (CCIEP). The hyperimmune polyclonal antisera were separately raised in rabbits against excretory/secretory (ES) antigens of both the flukes under study. An overall 70% buffalo faecal samples were tested positive for G. crumenifer and 75% for G. explanatum in Aligarh region. The ELISA results reflected higher infection intensity among individual buffaloes that was also observed at necropsy. Using the respective homologous hyperimmune antiserum, 55% buffaloes tested positive for G. crumenifer and 65% positive for G. explanatum in immunodot assay. Further, the faecal samples with high absorbance values in ELISA and strong immunodot reaction tested positive in CCIEP. The analysis of CCIEP result revealed two and one precipitin bands in G. crumenifer and G. explanatum respectively, indicating prominent antigenic differences in the coproantigens of these two parasites. Taken together, it is suggested that polyclonal antibodies could be conveniently used for the detection of coproantigens by ELISA and immunodot methods, particularly during the non-egg producing phase of the seasonally regulated reproductive cycle of the rumen amphistome G. crumenifer. It is concluded that the coproantigen detection is a good alternative over conventional method for the diagnosis of amphistomosis in livestock; however, further studies are required on a larger sample size of field buffaloes to augment the reproducibility of the present results.

Population structure and genetic variability in the Murrah dairy breed of water buffalo in Brazil accessed via pedigree analysis

The objective of this study was to use pedigree analysis to evaluate the population structure and genetic variability in the Murrah dairy breed of water buffalo (Bubalus bubalis) in Brazil. Pedigree analysis was performed on 5,061 animals born between 1972 and 2002. The effective number of founders (fe) was 60, representing 6.32 % of the potential number of founders. The effective number of ancestors (fa) was 36 and the genetic contribution of the 17 most influent ancestors explained 50 % of the genetic variability in the population. The ratio fe/fa (effective number of founders/effective number of ancestors), which expresses the effect of population bottlenecks, was 1.66. Completeness level for the whole pedigree was 76.8, 49.2, 27.7, and 12.8 % for, respectively, the first, second, third, and fourth known parental generations. The average inbreeding values for the whole analyzed pedigree and for inbreed animals were, respectively, 1.28 and 7.64 %. The average relatedness coefficient between individuals of the population was estimated to be 2.05 %-the highest individual coefficient was 10.31 %. The actual inbreeding and average relatedness coefficient are probably higher than estimated due to low levels of pedigree completeness. Moreover, the inbreeding coefficient increased with the addition of each generation to the pedigree, indicating that incomplete pedigrees tend to underestimate the level of inbreeding. Introduction of new sires with the lowest possible average relatedness coefficient and the use of appropriate mating strategies are recommended to keep inbreeding at acceptable levels and increase the genetic variability in this economically important species, which has relatively low numbers compared to other commercial cattle breeds. The inclusion of additional parameters, such as effective number of founders, effective number of ancestors, and fe/fa ratio, provides better resolution as compared to the inclusion of inbreeding coefficient and may help breeders and farmers adopt better precautionary measures against inbreeding depression and other deleterious genetic effects.

Serological and bacteriological responses of water buffalo (Bubalus bubalis) vaccinated with two doses of Brucella abortus strain RB51 vaccine

Thirty-two water buffalo (Bubalus bubalis) calves aged 6–10 months were used to evaluate serological responses to Brucella abortus strain RB51 (RB51) vaccination in a dose-response study and to compare the use of two selective media for the isolation of RB51. The animals were randomly divided into three treatment groups. Groups I-III received the recommended vaccine dose (RD) twice 4 weeks apart, RD twice 18 weeks apart and saline once, respectively. Lymph nodes were excised from the three groups and subjected to bacteriological examination to determine the frequency of detection of RB51. Pre- and post-vaccination blood samples were collected and tested for B. abortus antibodies using the buffered plate agglutination test (BPAT), complement fixation test (CFT), and dot-blot assay. Sera taken at all post-inoculation weeks (PIW) were negative for field strain B. abortus using the BPAT. Antibody responses to RB51 were demonstrated in all vaccinates but not in controls by CFT and dot-blot assay from 1 PIW up to 16 weeks following booster vaccination. The agreement for both assays was 80.7% and there was a linear interdependence with a Pearson's correlation coefficient value of 0.578. The frequency of isolation of RB51 from the two selective media used was not significantly different (P > 0.05).

Molecular mining of GGAA tagged transcripts and their expression in water buffalo Bubalus bubalis

Repeat sequences are involved in regulation of gene expression both at the transcriptional and translational level. In the mammalian genomes, tri- and tetranucleotide repeats like ATA, AATA, GGAA and GAAA have been associated with diseases. In silico analysis of (GGAA) 5 distribution across the species showed maximum number of this repeat in the mouse transcriptome compared to that in other species. Following this, we conducted minisatellite associated sequence amplification (MASA) to explore the buffalo's transcriptome using cDNA from different tissues and an oligo based on (GGAA) 5 repeats. MASA uncovered twenty six mRNA transcripts showing homology to known genes in the database. qPCR studies showed the highest expression of twelve transcripts in the spleen. A transcript, pLRC107 with its partial sequence of 203 nucleotides showed sequence variation at several positions in spleen as compared to other four tissues examined. Transcript pLRC100 was found to represent the partial coding sequence of Bos taurus HECT {(homologous to E6-associated protein (UBE3A) carboxyl-terminus domain) and RCC1 (CHC1)-like domain (RLD) 1}, mRNA. We ascertained full length coding sequence of HECT gene and localized the same on buffalo chromosome 10 employing FISH. This gene was found to be conserved across the species. Another gene LRP8 uncovered in the process showed copy number variation between buffalo males (4–9) and females (34–54). The MASA approach enabled us to identify several genes in Bubalus bubalis without screening an entire cDNA library. The highest expression of 12 mRNA transcripts in spleen suggests their likely involvement with immuno transaction. A comprehensive knowledge of the repeat tagged transcriptomes is envisaged to help in understanding their significance in genome organization and evolution forming rich basis of functional and comparative genomics.