Molecular detection of hemoplasmas and genetic diversity of Anaplasma marginale in a dairy water buffalo (Bubalus bubalis) farm from southeastern Brazil.

Isolation of verocytotoxin-producing Escherichia coli O157 from water buffaloes (Bubalus bubalis) in southern Italy

The swamp buffalo of Pampangan (swamp buffalo) is one of South Sumatran germplasm that has the potential to be developed as one of the livelihoods of the people in peat swamp land. Currently, its farming extends from the farm area of origin in Pampangan area to the development area in Riding Village, Pangkalan Lampam Subdistrict, Ogan Komering Ilir Regency. Its expansion is mostly located within production forest area which is adjacent to Wildlife Sanctuary (WS) of Padang Sugihan. In the last decade, its population has increased both by birth and displacement. This phenomenon may cause competition in obtaining food and trigger damage to Padang Sugihan WS ecosystem. In order to support the sustainability of its business and to maintain the preservation of Padang Sugihan WS ecosystem, the optimum management of buffalo farming is required. Biophysical analysis of optimum grazing area, optimization studies and livestock business analysis are the methods used in this study. The results show that the total area of grazing area outside Padang Sugihan WS area is 1,646 ha and optimum population per ha is 3 buffalos. Analysis of its business analysis showed that financial feasibility was feasible. They indicate that buffalo farm business is potential developed as an alternative to community livelihoods in peat lands. In order to maintain the preservation of forest ecosystems and sustainability of community livelihoods, it is necessary to improve the grazing pattern and management of grazing areas. Hence the fodder is fulfilled without damaging the area of Padang Sugihan WS. Introduction.

ClassyFarm is an integrated system for categorizing farms according to the risk assessment (RA) methodology. It is an Italian innovation aimed at improving the synergy between breeders and competent authorities to improve the safety and quality of food of animal origin. ClassyFarm gathers and processes data referred to in the following areas: biosecurity, animal welfare, health, and antimicrobial usage. It can be applied to livestock species, including water buffaloes. Upon request of the Italian Ministry of Health (IMH), the National Reference Centre on Water Buffalo Farming and Production Hygiene and Technologies (CreNbuf), in collaboration with the Italian Reference Centre for Animal Welfare (CreNBA), developed a RA-based checklist (CL) for the on-farm assessment of buffalo welfare and farm biosecurity level, included in the ClassyFarm system. The multiple- choice CL consists of 79 items. Each item is scored according to 3 categories: “insufficient”, “acceptable” and “excellent”. The assessment system for animal welfare includes non-animal-based (N-ABMs) and animal-based measures (ABMs). N-ABMs are divided into two macro-areas: Area A (32 items): “Management factors” and Area B (31 items): “Housing factors”. ABMs are assessed in Area C (17 items). Biosecurity was assessed using 15 indicators. The CL has been tested in 102 farms, with an average size of 412 heads. On average, the overall welfare value was 61.55% (on a scale from 0 to 100%), and the average biosecurity score was 43.31%. The statistical analysis was performed by Spearman Rank correlation coefficient using GraphPad Prism 8.0.1. (GraphPad Software, San Diego, CA, USA). The two variables were positively correlated (Spearman’s Rho=0.501; p<0.001). The average welfare values of the specific areas were: A, 61.76%; B, 42.17%; C, 70.43%. At least one potential legislative non-compliance was recorded in 39.80% of the farms. The highest correlations between the different areas of well-being and biose-curity are found for the management areas, highlighting how management is essential to ensure both the levels of well-being and those of biosecurity. These CLs represent a functional and effective tool to assign animal welfare and biosecurity indexes to farms, improving farm management and housing conditions, giving answers to consumers, and adding value to farmers’ good practices. The IMH is promoting the application of this system at European and international levels.

Brazil has the largest buffalo herd outside the Asian continent, with more than 1.5 million head. As buffalo farming has become an economically important activity, studies on the occurrence of pathogens shared between cattle and buffalo are necessary. The aim of this study was to investigate the molecular occurrence of piroplasmids and bacterial agents (Anaplasmataceae and Coxiellaceae) potentially transmitted by vectors in buffaloes in southeastern Brazil using molecular methods. DNA was extracted from blood samples of 81 buffaloes, 165 Rhipicephalus microplus ticks, and 92 Haematopinus tuberculatus lice collected from buffaloes in the municipality of Passos, state of Minas Gerais, Brazil. These samples were subjected to conventional PCR (cPCR) assays based on three endogenous genes, namely gapdh from mammals, 16S rRNA from ticks, and cox-1 from lice. Positive samples from these PCR assays were then submitted to a real-time PCR (qPCR) assay targeting IS1111 of Coxiella burnetii, cPCR for Ehrlichia spp. (dsb gene), and nested-PCR assays targeting the 18S rRNA gene of Piroplasmida, sbp-2 of Babesia bovis, and rap-1α of Babesia bigemina. All buffalo and ectoparasite DNA samples tested positive in the PCR assays for the endogenous genes. All samples were negative in the qPCR for C. burnetii and nested PCR for B. bigemina (rap-1α). For Ehrlichia spp., 6/257 (2.3%) ectoparasites (five ticks and one louse) tested positive, with BLASTn analysis showing 88.04% to 100% identity with Ehrlichia minasensis. In the nPCR for piroplasmids (18S rRNA), 8/81 (9.9%) buffaloes and one tick (0.7%) tested positive, with BLASTn analysis showing > 99.9% identity with B. bovis and B. bigemina. In the nPCR for B. bovis (sbp-2), 1/81 buffalo (1.2%) was positive. This study demonstrated, for the first time, the occurrence and co-infection of tick-borne hemoparasites in buffaloes and associated ectoparasites in southeastern Brazil. The lack of positivity for E. minasensis and low molecular occurrence for B. bovis and B. bigemina among buffaloes sampled may be associated with the higher resistance of this ruminant species to R. microplus ticks and vectored pathogens.

Buffalo breeding is common in Southern Italy. Dystocia compromises dam's and newborn health and welfare. Difficult parturition could be solved through prompt calving assistance, even if the identification of the beginning of delivery is challenging. Herein, we aimed to evaluate a remote calving alarm system in 15 Mediterranean buffalo heifers. An intravaginal probe was placed close to the external cervical os once premonitory signs of delivery were observed. No vaginal discharge nor signs of discomfort were notified in the days following the insertion of the probe. Heifers calved from 48 to 72hr after the alarm was activated. The system correctly warned the farm personnel at the beginning of stage II of parturition, except for 2 cases. In the former, the intravaginal probe was expelled but the poor carrier network coverage negatively affected phone's signal quality; in the latter, recurrent vaginal prolapse was responsible for non-retention of the probe. Overall median expulsive phase was 68±8min, while the expulsion of a female calf took 54±22.0min and 90±34.0min in males, with significant difference (p=.02). Deliveries were homogeneously distributed across a 24-hr interval. No retention of foetal membranes nor metritis was identified at postpartum clinical examination. The calving alarm system used in this work was well tolerated in buffalo heifers. The introduction of smart technology in buffalo farming could contribute to the overall farm net return by reducing calf losses, especially for calves born from sexed-sorted semen, and by increasing animal welfare through quick resolution of dystocia. Further studies will be necessary to evaluate the net return in buffalo farms which will implement a remote calving alarm system on a wider population.

The purpose of this study was to analyse the socio-economic structure of water buffalo farming in the province of Muş, in Turkey. The stratified sampling method was used to calculate sample size of buffalo farms in the Central, Korkut, and Hasköy districts of Muş province, where buffalo farming is widespread. Data were collected from the 94 farms by surveys in the 2013 production period. A single budget analysis method was used to calculate production cost and profit for water buffaloes. Plant production constituted 37.85% of the gross production value, while animal production accounted for 62.15%. The biggest share in the gross production value derived from water buffalo farming (45.71%). Fixed and variable costs were 51.44% and 48.56% of the production cost (USD11691.06), respectively. The largest part of the variable cost was the feed cost (75.81%). The cost of per kilogram buffalo milk in the region was calculated as USD0.64. Consequently, it is important to ensure the continuation of breeding studies to increase milk yield, giving information to farmers about modern techniques, developing policies to increase the scale of the farms, and implementing regional and national policies to increase awareness of buffalo milk and milk products.

Buffalo farming is a dairy enterprise emerging in Italy and other European and non-European countries due to the progressive saturation of the dairy market, which requires product diversification. As a response to the increasing demand for dairy buffalo products, most farms are undertaking a progressive intensification of rearing techniques. In particular, feeding rely on different rations according to the physiological stage of the animals. Buffalo heifers are generally offered a total mixed ration based on corn silage and ryegrass hay, which require high amounts of water, chemical fertilisers and fossil fuel. Additionally, lactating buffaloes receive concentrates with an even higher environmental impact, whereas more fibrous feeds are used in the dry period. Although the emission factors for enteric fermentation and manure management in buffaloes appear to be 38.4 % and 20.5 % less than in dairy cows, respectively, the contribution of dairy buffalo farms to global warming has been estimated as higher than that produced by dairy cow farms. Therefore, studies are needed to assess the general environmental impact of this species and identify alternative environmental friendly rearing techniques.

Water buffaloes have morphological and behavioral characteristics for efficient thermoregulation. However, their health, welfare, and productive performance can be affected by GW. The objective of this review was to analyze the adverse effects of GW on the productive behavior and health of water buffaloes. The physiological, morphological, and behavioral characteristics of the species were discussed to understand the impact of climate change and extreme meteorological events on buffaloes' thermoregulation. In addition, management strategies in buffalo farms, as well as the use of infrared thermography as a method to recognize heat stress in water buffaloes, were addressed. We concluded that heat stress causes a change in energy mobilization to restore animal homeostasis. Preventing hyperthermia limits the physiological, endocrine, and behavioral changes so that they return to thermoneutrality. The use of fans, sprinklers, foggers, and natural sources of water are appropriate additions to current buffalo facilities, and infrared thermography could be used to monitor the thermal states of water buffaloes.

Toxoplasmosis is a worldwide widespread zoonotic infection caused by the intracellular protozoan Toxoplasma gondii. This infection is considered one of the most important food-borne parasitic zoonoses globally. In addition to its public health impact, toxoplasmosis also has important veterinary implications, as it causes abortions or congenital malformations with negative economic impacts. Several serological studies on toxoplasmosis in water buffaloes (Bubalus bubalis) are fragmentary, dated, consider a low number of animals compared to the present population and do not relate the epidemiological update in an area of the Mediterranean. For this reason, the objectives of this study were to define the seroprevalence of T. gondii infection in water buffaloes and to assess the risk factors associated with infection in water buffalo farms. A total of 184 dairy buffalo farms were randomly selected upon arrival of blood samples collected during state prophylaxis and analyzed with an indirect ELISA kit (ID Screen®, Indirect Toxoplasmosis Multi-Species, IDVET, France), according to the manufacturer›s instructions. The random selection of farms allowed us to consider different areas of the Campania region where the highest concentration of buffalo herds exists. For the risk assessment study, a specially designed questionnaire was administered that considered several factors (number of animals, presence of cats and rodent control measures) as well as presence of clinical signs such as abortions in adult buffaloes. Of a total of 13,649 animals analyzed, 2,693 tested positive for T. gondii (19.7%; 95% Confidence Interval [CI] = 19.7-24.6%), and out of 184 buffalo farms 179 tested positive (97.3%; 95% CI = 93.8- 98.8%). Of the T. gondii positive farms, 97% had cats on the farm and 85% did not implement rodent control measures. These findings indicate that these two risk factors are the most prevalent and need to be monitored for infection control on buffalo farms.

As part of the agricultural sector, people in the Sijunjung district also make the livestock subsector part of the family’s economic source, where the type of livestock that has been cultivated for a long time is buffalo. Sijunjung is one of the areas in the Sijunjung district which has the potential to develop buffalo farms because it has the highest livestock population. This study aims to determine the model of buffalo livestock raising, and the income of buffalo farmers in Sijunjung District and to analyze the factors that affect the income of buffalo farming in Sijunjung District. This research method uses a survey method by interviewing 35 respondents. Data analysis used descriptive statistical analysis methods and inferential statistics using multiple linear regression analysis. The results showed that most of the farmers (40%) had buffalo in the range of 1-5 heads. Buffalo livestock farming is only used as a side business, where only 17.1% make buffalo farming their main business. Furthermore, the status of livestock ownership in general (65.7%) is a profit-sharing system that in West Sumatra is known as seduaan. The buffalo-rearing system is semi-intensive. The average income of farmers from buffalo farming for one year is Rp. 8,618,669, -. The factor of farm scale, age of the farmer, and the number of calves showed a significant effect on the income of the buffalo farming, while the factors of the number of buffaloes, education, experience, and fixed costs did not significantly affect the income of the buffalo farming.

A cross-sectional study was conducted to estimate the prevalence of intramammary infection (IMI) associated bacteria and to identify risk factors for pathogen group-specific IMI in water buffalo in Bangladesh. A California Mastitis Test (CMT) and bacteriological cultures were performed on 1,374 quarter milk samples collected from 763 water buffalo from 244 buffalo farms in nine districts in Bangladesh. Quarter, buffalo, and farm-related data were obtained through questionnaires and visual observations. A total of 618 quarter samples were found to be culture positive. Non-aureus staphylococci were the predominant IMI-associated bacterial species, and Staphylococcus (S.) chromogenes, S. hyicus, and S. epidermidis were the most common bacteria found. The proportion of non-aureus staphylococci or Mammaliicoccus sciuri (NASM), S. aureus, and other bacterial species identified in the buffalo quarter samples varied between buffalo farms. Therefore, different management practices, buffalo breeding factors, and nutrition were considered and further analyzed when estimating the IMI odds ratio (OR). The odds of IMI by any pathogen (OR: 1.8) or by NASM (OR: 2.2) was high in buffalo herds with poor milking hygiene. Poor cleanliness of the hind quarters had a high odds of IMI caused by any pathogen (OR: 2.0) or NASM (OR: 1.9). Twice daily milking (OR: 3.1) and farms with buffalo purchased from another herd (OR: 2.0) were associated with IMI by any pathogen. Asymmetrical udders were associated with IMI-caused by any bacteria (OR: 1.7). A poor body condition score showed higher odds of IMI by any pathogen (OR: 1.4) or by NASM (OR: 1.7). This study shows that the prevalence of IMI in water buffalo was high and varied between farms. In accordance with the literature, our data highlight that IMI can be partly controlled through better farm management, primarily by improving hygiene, milking management, breeding, and nutrition.

First molecular detection of Mycoplasma wenyonii and the ectoparasite biodiversity in dairy water buffalo and cattle in Bohol, Philippines

The objective of this study was to determine the effect of a short five-day (5 d) progesterone-based timed artificial insemination (TAI) protocol on the time of ovulation and fertility in dairy water buffaloes. In experiment 1, 30 dairy buffaloes were subjected to one of two estrous synchronization protocols. Females in the control group (n=17) were administered 2 mg of estradiol benzoate (EB), and an intravaginal progesterone-releasing device (CIDR) was inserted on day -12. On day -3, CIDR inserts were removed and all cows were treated with 150 ?g of D-cloprostenol (a PGF2?-analogue). Forty-eight hours after CIDR removal, all cows received 100 µg of gonadotropin-releasing hormone (GnRH). Females assigned to the short 5 d group (n=13) received treatment similar to the control group, except that the CIDR device was inserted on day -8, and the EB treatment was eliminated. In experiment 2, 51 dairy buffaloes were separated into control (n=27) and short 5 d (n=24) groups that received the same treatment as described for cows in experiment 1, except that all females were inseminated 16 to 18 h after GnRH injection. The use of a short 5 d progesterone-based TAI protocol with or without treatment with EB on day 0 did not affect the time of ovulation and pregnancy per artificial insemination in lactating water buffaloes. Thus, we suggest that the short 5 d TAI protocol may be successfully used as an alternative to induce ovulation in dairy buffaloes, with the particular advantages of avoiding EB treatment and a shorter interval from the beginning of the protocol to TAI.

Insulin-like growth factor-1 (IGF-1) gene plays an important role in the endocrine system of animals by regulating nutrient metabolism, growth, and milk production. There have been extensive molecular genetics research studies on cattle but less studies have focused on buffalo (Bubalus bubalis). This study aimed to confirm the association of IGF-1 gene in swamp or river buffalo (B. bubalis spp.) with growth and milk production traits. DNA samples were obtained from 12 buffalos (eight swamp buffalo and four river buffalo). One Bali cattle (Bos javanicus) was included as an outgroup (control). The eight swamp buffalo originated from East Nusa Tenggara (n = 1), Baluran, East Java (n = 4), and Banyuwangi, East Java (n = 3) , while the four river buffalo originated from Sei Putih, Medan of North Sumatera. All DNA samples were amplified using an IGF-1 primer for 30 cycles, and amplicons were visualized on 1% agarose gel. Five of the 13 samples were sequenced to determine nucleotide sequence variation between the swamp and river buffalo. The results revealed that the size (225–231 bp) of all the fragments was in in accordance with that of IGF-1. There was not found genetic variation among the buffalo samples. The results indicate that buffalo samples bear growth and milk production traits.

First molecular detection and characterization of tick-borne pathogens in water buffaloes in Bohol, Philippines