Bubalus Research Articles

A Biodiversity Conservation Area Using Holistic and Modular Architecture

Green architecture and engineering aim to minimize the use of available resources by maintaining and protecting the ecosystem and optimizing its benefits to humans and the ecology. The rich biodiversity of Mindoro Island, Philippines has always been on the pipeline project of the province. However, strategies for maximizing the benefits of a small area imposed challenges and hindered many conservation projects. This study utilized modular architecture, biophilia, and biomimesis as design concepts. The design implemented the use of inverted pyramid-shaped modules supported by pilotis to lessen the interaction of the construction project to the proposed site. Five modules are built accordingly and strategically connected by a modular bridge for accessibility. The body of the Mindoro dwarf buffalo is represented by the container van used as the mainframe of the modules; the legs are represented by the pilotis used as structural columns, and the body-built is represented by the inverted pyramid-shaped single-pile structures. The administration building was also inspired by the sturdy built of Mindoro Dwarf Buffalo (Bubalus mindorensis) and designed to cater the administrative functions and activities, while the proposed shops and commercial areas are designed to boost the local economy.

Draft genome of the lowland anoa (Bubalus depressicornis) and comparison with buffalo genome assemblies (Bovidae, Bubalina).

Genomic data for wild species of the genus Bubalus (Asian buffaloes) are still lacking while several whole genomes are currently available for domestic water buffaloes. To address this, we sequenced the genome of a wild endangered dwarf buffalo, the lowland anoa (Bubalus depressicornis), produced a draft genome assembly and made comparison to published buffalo genomes. The lowland anoa genome assembly was 2.56 Gbp long and contained 103,135 contigs, the longest contig being 337.39 kbp long. N50 and L50 values were 38.73 and 19.83 kbp, respectively, mean coverage was 44× and GC content was 41.74%. Two strategies were adopted to evaluate genome completeness: (1) determination of genomic features with de novo and homology-based predictions using annotations of chromosome-level genome assembly of the river buffalo and (2) employment of benchmarking against universal single-copy orthologs (BUSCO). Homology-based predictions identified 94.51% complete and 3.65% partial genomic features. De novo gene predictions identified 32,393 genes, representing 97.14% of the reference's annotated genes, whilst BUSCO search against the mammalian orthologs database identified 71.1% complete, 11.7% fragmented, and 17.2% missing orthologs, indicating a good level of completeness for downstream analyses. Repeat analyses indicated that the lowland anoa genome contains 42.12% of repetitive regions. The genome assembly of the lowland anoa is expected to contribute to comparative genome analyses among bovid species.

Polymorphism in inhibin alpha gene and its association with semen quality traits in Murrah bulls

An investigation was conducted to detect single nucleotide polymorphism in Inhibin alpha gene (INHA) and to analyse the association between the observed polymorphisms with semen quality traits in Murrah buffalo bulls. Genomic DNA was isolated from blood samples of 103 bulls, the amplified PCR products were further subjected to polyacrylamide gel electrophoresis. Band patterns were characterised by the number of bands, mobility shifts and scored manually. Representative PCR products giving unique SSCP patterns were custom sequenced using automated ABI DNA Sequencer to confirm the mobility shift in each pattern. Sequence data were analysed using DNA Baser and Bio edit software Clustal omega multiple alignments for detecting single nucleotide polymorphisms in comparison to observed sequence with the Bubalus sbubalis INHA gene reference sequence. INHA gene, exon1 was monomorphic, while exon 2 was found to be polymorphic. For simple interpretation of SSCP patterns, INHA exon 2 was divided into two fragments. The first fragment of INHA exon 2 (INHA exon 2 FR1) revealed a transition at the C267119T position and a transversion at the G267033T position in the coding region in comparison to Bubalus bubalis reference sequence. The second fragment of INHA exon 2 (INHA exon 2 FR2) exhibited one transition at T266510C position in comparison to the Bubalus bubalis reference sequence. The SSCP variants of the first fragment of INHA exon2 had a significant effect on semen volume per ejaculate (P<0.05) and sperm concentration (P<0.01). The SSCP variants of the second fragment of INHA exon 2 had a significant effect on sperm concentration (P<0.01) and functional membrane integrity (P<0.05). The observed association between SSCP variants in INHA gene with semen volume per ejaculate, sperm concentration and functional membrane integrity suggests the possibility of using these genetic variants in INHA gene as prognostic markers for selecting Murrah bulls with high semen quality traits.

Una leche bubalina rica en ácidos grasos trans incrementó la colesterolemia y tendió a agravar la ateroesclerosis en conejos

&lt;p&gt;Búfalas suplementadas con una mezcla de aceites de girasol + pescado pueden reducir en la leche los niveles de ácidos grasos saturados (AGS), considerados hipercolesterolémicos y pro-aterogénicos, e incrementar los ácidos grasos trans-11 18:1 y cis-9,trans-11 18:2, con propiedades anti-aterogénicas. Este trabajo comparó los efectos del consumo de dos leches bubalinas, con baja y alta relación de ácidos grasos trans (AGt)/AGS, sobre el lipidograma y el desarrollo de ateroesclerosis inducidos por colesterol en conejos. Veinte conejos neozelandeses, machos, fueron aleatoriamente separados en un grupo control (n=10) que recibió leche con baja relación AGt/AGS (5,3% de AGt y 67,12% de AGS) obtenida de búfalas en sistema pastoril; y un grupo alto trans (n=10) que recibió leche con alta relación AGt/AGS (25,84% de AGt y 45,89% de AGS) obtenida de búfalas con suplementación lipídica en su dieta. La experiencia duró 75 días y, a partir del día 15, todos los conejos fueron desafiados con 0,93 g de colesterol/día, vía oral, a fin de inducir lesiones ateroescleróticas. Los conejos consumieron de manera voluntaria 9293,13 ml y 9930 ml de leche con baja y alta relación AGt/AGS, respectivamente (p 0,404). Los conejos que consumieron leche con alta relación AGt/AGS registraron un leve incremento significativo (p 0,049) del colesterol total (6,08 g/l), cuando se comparó con el grupo control (5,58 g/l). No se detectaron diferencias entre grupos en el colesterol LDL (p 0,073), colesterol HDL (p 0,078) y triglicéridos (p 0,174). Las lesiones ateroescleróticas fueron más extensas en los conejos que consumieron leche con alta relación AGt/AGS, aunque sin significación estadística (p&amp;gt;0,05) cuando se comparó el control. En conclusión, la leche bubalina obtenida con la suplementación lipídica referida en este estudio, incrementó la colesterolemia y tendió a agravar la ateroesclerosis en conejos. Dichos efectos, posiblemente estén relacionados con el alto contenido de AGt de esta leche.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;

Anoa, dwarf buffalo from Sulawesi, Indonesia: Identification based on DNA barcode

Priyono DS, Solihin DD, Farajallah A, Arini DID. 2018. Anoa, dwarf buffalo from Sulawesi, Indonesia: Identification based on DNA barcode. Biodiversitas 19: 1985-1992. Anoa is an endangered endemic species in Sulawesi. The controversial issue of anoa conservation until now is the taxonomic status of lowland and mountain anoa. This study aims to test the ability of DNA barcoding techniques to identify the taxonomy between mountain anoa and lowland anoa. A 681bp fragment of cytochrome oxidase subunit 1 (COI) gene was obtained and used to solve the molecular taxonomic problem and to resolve the phylogenetic relationships of the two types of anoa. Our results showed that the DNA barcode is useful in assigning the taxonomic position of anoa. In the phylogenetic tree, we found that the two types of anoa were in separate clades. We also found that based on the Kimura-2 parameter (K2P), the genetic distance between the two types of anoa showed higher values (3.4%) than the threshold of the separating species level. We, therefore, proposed that the binomial nomenclature for both types of lowland and mountain anoa are respectively Bubalus depressicornis and Bubalus quarlesi. We suggest that the use of DNA barcode techniques in anoa taxonomic studies and their implementation will be useful in conservation management

Detection of Cryptosporidium parvum DNA in fecal samples of infected cattle (Bos indicus) and water buffaloes (Bubalus bubalis) in the Philippines using loop mediated isothermal amplification method

Fecal DNA samples from 17 cattle and 38 water buffaloes found to be infected with Cryptosporidium oocysts using Kinyoun acid fast stain from a previous study, were subjected to loop-mediated isothermal amplification (LAMP) assay using specific primers for Cryptosporidium parvum (C. parvum) from three municipalities, Maria, Baler and San Luis of the province of Aurora in the Philippines. Results of the fecalysis using Kinyoun acid fast stain and LAMP assay were compared with the PCR results of the examined farmer/owner who raised these animals to determine the possible zoonoses of C. parvum between the farmers and their animals. Using LAMP assay, only 41% (7/17) were positive in cattle and 76% (29/38) in water buffaloes. Out of the seven LAMP positive cases in cattle, 86% (6/7) came from Maria and 14% (1/7) from Baler. Out of 29 LAMP positive cases in water buffaloes, 62% (18/29) came from Maria, 24% (7/29) from Baler and 14% (4/29) from San Luis. Comparing with the earlier results for probable zoonoses of C. parvum between the farmers and their animal was determined. Eight farmers that were positive in PCR and with their water buffaloes, positive in LAMP assay were detected to have C. parvum. Only one farmer with his cattle was detected positive of Cryptosporidium spp. in PCR, however, it was negative in LAMP assay hence, a non-parvum species might infected the farmer and the animal.

Identification of microRNAs in corpus luteum of pregnancy in buffalo (Bubalus bubalis) by deep sequencing.

This study was aimed to identify miRNAs of corpus luteum (CL) in buffaloes during pregnancy. For this study, CL (n=2) were collected from gravid uteri of buffalo and RNA was isolated. Following this, the purity and integrity of RNA was checked and used for deep sequencing using Illumina Hiseq 2500 platform. The reads' quality was checked prior to in silico analyses viz. identification of conserved, novel and target of miRNAs. In this study, out of identified miRNAs (3018), 3013 were known and 5 were novel miRNAs on alignment with reference genomes. In addition, prediction of putative target genes for identified abundant miRNAs revealed several genes viz. HOX, KLF4, NCOR2, CDKN2Z, MAPK7, COX2, PPARA, PTEN, ASS3A, ELK1, CASP3, BCL211, MCL1, CCND2, Cyclin A2 and CDC25A during early pregnancy in buffalo. These predicted target genes have been associated with various cellular house-keeping processes including apoptosis. In conclusion, this study reports the identification of conserved and novel microRNAs (miRNAs) in CL during pregnancy in buffalo by deep sequencing.

The Histological Structure of the Ileal Peyers Patches of the Egyptian Water Buffalo (Bos Bubalus)

The water buffalo represents an important source of animal products in Egypt and characterized by high resistance to several diseases and parasites. The present study carried out on the Egyptian water buffaloes of different ages (from 50 days to 8 years old buffaloes) to illustrate the structure of ileal Payer's patches with age. The specimens of ileum were fixed and processed for light and electrone microscopy The Payer's patches (PPs) formed of elongated dome regions flanked by intestinal villi. These domes were short, wide villi. The domes were consisted of lymphoid follicles covered with a typical dome- associated epithelium of enterocytes and M cells without any goblet cells in young and adult ages. The M cells showed variable appearance depending on the functional status. The lymphoid follicles had a clear germinal center. In conclusion Payer's patches (PPs) were still active until the age of 5 years old and then begin to be inactive at 8 years old we think there is a long period of activity which express the tolerance to many diseases.

Evaluating a murine model of endometritis using uterine isolates of Escherichia coli from postpartum buffalo.

Ascending infection of the uterus with Gram-negative bacteria is responsible for postpartum endometritis in cattle and buffalo and can adversely affect fertility. Development of a laboratory animal model for bovine endometritis would facilitate the understanding of the pathogenesis as it is difficult to conduct controlled experimentation in the native host. In the present study, 30 virgin Swiss Albino mice (5-8 weeks old) were used to evaluate the pathogenic potential of Escherichia coli, isolated from the normally calved postpartum buffalo to induce endometritis. Mice in the diestrus phase of the estrous cycle were randomly allotted to one of the following four intravaginal inoculation (100 μL) treatments: EG (experimental group)-1: sterile normal saline; EG-2, -3 and -4: E. coli@ 1.5 × 104, 105 and 106 CFU/ml, respectively. The animals were then scarified 36 h post-inoculation to study gross and microscopical lesions. Gross changes were confined to EG-4. Acute endometritis was recorded in 50% of the EG-3 and 66.7% of the EG-4. The rate of acute endometritis development was significantly higher in EG-4 (P<0.05) as compared to the other groups. The present study demonstrated that the animal model for bubaline endometritis can be developed in mice by intravaginal inoculation of E.coli@ 1.5 × 106 CFU/ml at diestrus. Ease of intravaginal inoculation, apparent absence of systemic involvement and high infective rate are the advantages of the model over other studies.

Humoral immune response of buffalo heifers (Bubalus bubalis) vaccinated with B19 strain of Brucella abortus

RESUMO: A brucelose se encontra distribuída mundialmente. No Brasil apresenta maior prevalência para a espécie Brucella abortus que apesar de ser endêmica, encontra-se distribuída de forma heterogênea entre as diferentes regiões do Brasil. O presente estudo teve por objetivo avaliar a resposta imune humoral de bezerras bubalinas criadas na Mesorregião do Médio Amazonas, vacinadas com a cepa B19 de B. abortus na idade preconizada pelo Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT). Os animais selecionados foram 36 fêmeas (grupo teste) e 6 machos (grupo controle) com idades entre 3-8 meses. No dia 0 foi realizada a colheita de sangue em todos os animais, em seguida as fêmeas receberam vacina comercial com amostra B19 em dose padrão, com posteriores coletas de sangue ate aos 390 dias. Os soros sanguíneos foram avaliados nas provas de soro aglutinação: Antígeno Acidificado Tamponado - AAT e 2- Mercaptoetanol - 2ME. Os resultados para todas as amostras coletas no dia 0 apresentaram resultado negativo na reação. Aos 30 e 60 dias todas as fêmeas apresentaram 100% de reação na prova AAT e titulação de 200, iniciando o declínio da reação aos 90 dias nas duas provas. Durante o estudo um animal se mante reativo nas duas provas até 360 dias e, somente aos 390 dias 100% das fêmeas obtiveram reação negativa nas duas provas, durante todo o estudo os machos foram não reativos. A vacinação mostrou-se eficaz para a imunização, sendo uma ferramenta importante na profilaxia da brucelose na espécie estudada, bem como as provas aplicadas para o diagnostico dessa enfermidade em búfalos a nível regional.

Comparative efficacy of E-17β and GnRH administration on day 0 of a controlled internal drug release (CIDR) based protocol on synchrony of wave emergence, ovulation and conception rates in Murrah buffalos (Bubalus bubalis).

The study was undertaken to compare the efficacy of an estradiol-17β + CIDR based protocol with the conventional ovsynch + CIDR based protocol for synchrony of wave emergence and ovulation in Murrah buffalos. In group I (n=25), on day 0 (beginning of experiment), buffaloes were administered a CIDR device (1.38 g P4) and concurrently received 1.5 mg E-17β. On day 9, the CIDR was removed and a prostaglandin (PG) F2α analogue (500 µg) was administered. On day 11, buffaloes were administered a gonadotropin releasing hormone (GnRH) analogue (20 µg) and inseminated twice at 12 h and 24 h following GnRH injections. Group II (n=25) protocol was based on an ovsynch regimen plus CIDR for 7 days followed by double insemination at induced estrus. Group III (n=10) served as control and was not given any hormone on day 0 of the protocol. In groups I, II and III, the duration of new follicular wave emergences were observed on days 4.22 ± 0.12, 3.12 ± 0.33 and 5.14 ± 0.42, respectively. In group I, synchrony of wave emergence was more and the diameter of pre-ovulatory follicles was larger (P<0.05) compared to groups II and III. The first service conception rate (FSCR) was higher (P<0.05) in group I while ovulation rates were not different between groups I and II. In conclusion, more synchrony of wave emergence, larger diameter of dominant follicles and higher first service conception rate was observed following the E-17β + CIDR based protocol in buffalos.

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.

Hemal Node of Egyptian Water Buffalos (Bos Bubalus): It’s Role in Erythrophagocytosis

Hemal nodes were obtained from 12 clinically healthy male buffalos aged 2-3 years, their average weight is 500-600kg. Electron microscopy revealed macrophages engulfing intact erythrocytes in vacuoles, or only parts of erythrocytes appeared in the vacuoles. Some electron dense material appeared in the erythrocyte as a lysosomal degrada- tion. Also myeline like figures and residual bodies were seen in macrophages cytoplasm.

IN VITRO EVALUATION OF BUBALINE ACELLULAR SMALL INTESTINAL MATRIX

In this study, a decellularization process was employed to remove the cellular components from small intestine of buffalo ( Bubalus bubalis ) using 2% ionic biological detergent. The acellular matrix then was cross linked with 0.6% glutaraldehyde (GA), 1% 1, 4-butanediol diglycidyl ether (BDDGE) and 1% 1- ethyl- 3 - (3 - dimethylaminopropyl) carbodiimide (EDC). The physical properties of cross linked samples were compared, in vitro , to those of the acellular counterparts on the basis of gross observations, degradation tests, free protein contents, free amino group contents, fixation index, free hydroxyproline contents determination, moisture percent and SDS-PAGE analysis. GA treated tissue showed highest resistance to nonenzymatic, enzymatic degradation and significant reduction (P<0.05) of free protein contents, free amino groups contents and moisture percentage. SDS-PAGE analysis revealed that GA treated acellular small intestinal matrix expressed highest cross-linking, followed by EDC, and BDDGE treated tissues.

Comparison of somatic cell count, California mastitis test, chloride test and rennet coagulation time with bacterial culture examination to detect subclinical mastitis in riverine buffalo (Bubalus bubalis)

The study investigates the etio-prevalence of bubaline subclinical mastitis (SCM) with reference to relationship between somatic cell counts (SCC), California mastitis test (CMT), rennet coagulation time (RCT) and chloride test with bacterial cultural examination. Each test was conducted by standardized protocol. The prevalence of subclinical mastitis (SCM), latent infected quarters and non-specific infected quarters were found to be 17.83, 2.89 and 3.06%, respectively, when partitioned on the basis of International Dairy Federation criteria for SCC. Staphylococcus spp. (44.70%) and Streptococcus spp. (34.20%) were the most prevalent etiological agents isolated in this study. The mean values of SCC and RCT were compared separately in healthy and SCM milk infected by different etiological agents by Duncan multiple range test. Mean comparison for these values for healthy and SCM milk was done by t-test. The elevation recorded for SCC and RCT was highly significant (p<0.01) irrespective of the bacterial agents causing SCM. After calculating the percent sensitivity, specificity and accuracy, predictive values and likelihood ratios, it can be accomplished that SCC was mostly in agreement with bacterial culture examination. It is recommended that it is better to cross match the result of SCC with RCT and CMT. Chloride test is not suggested to diagnose SCM in riverine buffaloes. Key words: Bacterial culture, buffalo, California mastitis test, chloride test, rennet coagulation time, somatic cell count, subclinical mastitis.

Male-specific repeats in wild Bovidae

In this study, we investigated repetitive sequences localized on Y chromosomes. Repetitive DNA sequences represent a substantial part of the eukaryotic genome and, among them, a large portion comprises sequences repeated in tandem. Efficient and rapid isolation of repeat units is possible due to a laser microdissection technique used for Y chromosome separation, followed by polymerase chain reaction (PCR), cloning, and sequence analysis. We applied the derived repeat units to members of nine tribes within the Bovidae. Apart from the Y chromosomes of Bos taurus and Bubalus bubalis, where we used known sequences of repetition, the derived sequences were used as probes for fluorescent in situ cross-hybridization to members of the nine tribes of the Bovidae. We investigated the distribution of repeat units within the tribes and their localization on the Y chromosome. Sharing of sequence variants would indicate common descent, while the rapid horizontal evolution should allow discrimination between closely related species or subspecies.

45. Carbon-13 and Nitrogen-15 Turnover in Serum of Bubaline Donors of Biological Material for Medical Use

Univ Estadual Paulista, Ctr Estudos Venenos & Anim Peconhentos CEVAP, Fac Med, Botucatu, SP, Brazil

The parthenogenetic development of buffalo (Bubalus bubalis) oocytes after chemical stimulation

In buffalo the overall in vitro embryo production efficiency is lower than cattle, mainly due to the lower cleavage rate (Gasparrini, 2002). In fact in our experience, comparable embryo yields are obtained in the two species only in relation to the cleaved oocytes. It is thought that a major limiting factor is the quality of the frozen semen. It has been demonstrated that freezing results in acrosomal damage, leakage of enzymes, alterations in ionic strength and pH, complete withdrawal of the hydration shell of protein in the solution and loss of motility (Meur et al., 1988). On the other hand we cannot role out that the lower cleavage rate is related to the failure in the accomplishment of oocyte competence after maturation. In fact although nuclear maturation is successfully achieved in our current system

Isolation of Mycoplasma from a buffalo (Bubalus bubalis) with mastitis.

Summary From the milk of a buffalo Mycoplasma were isolated and identified as M. laidlawii. Zusammenfassung Isolierung von Mykoplasmen aus einem Buffel (Bubalus bubalis) mit Mastitis Aus dem Milchdrusensekret eines Buffels wurden Mykoplasmen isoliert und als M. laidlawii identifiziert. Resumen Aislamiento de Mycoplasma de una bufala (Bubalus bubalis) con mastitis De la secrecion de la glandula mamaria de una bufala se aislaron mycoplasmas, identificandose como M. laidlawii.

Histology of hemal nodes of the water buffalo (Bos bubalus)

Hemal nodes are independent lymphoid organs found in various mammals but are ignored by most immunologists. They seem to play a role in defense against blood-borne infections in some species. The structure of the hemal node has been described in various species but, so far, not in the water buffalo. Specimens were obtained from ten clinically healthy male animals (five calves: 2-3 months old; five bulls: 2-8 years old). Six hemal nodes were obtained from each animal from the mesenteric and perirectal region. The samples were studied by light and transmission electron microscopy. The hemal nodes are bean-shaped or spherical, with one hilus through which the hilus arteries and nerves enter the node and from which veins and lymphatics leave it. The buffalo hemal node has a thin capsule of connective tissue and a few smooth muscle cells. Trabeculae extend from the capsule partially dividing the parenchyma. Subcapsular and trabecular blood sinuses are present. The parenchyma is composed of irregular lymphoid cords rich in erythrocytes, macrophages, and plasma cells and is separated by blood sinuses of variable size engorged with blood. These blood sinuses drain into the trabecular sinuses and then into the subcapsular sinus. In calves, the size of the lymphoid cords is larger than that in adult bulls. Buffalo hemal nodes can be classified as typical hemal nodes, because they are definitely different from hemolymph nodes in other species. They may play a role in filtering the blood.