Tropical tanniniferous legumes used as an option to mitigate sheep enteric methane emission

This study presents the first results from Brazil using SF₆ tracer technique adapted from cattle to evaluate the capability of condensed tannin (CT) present in three tropical legume forages, Leucaena leucocephala (LEU), Styzolobium aterrimum (STA), and Mimosa caesalpiniaefolia Benth (MIM) to reduce enteric CH₄ production in Santa Inês sheep. Twelve male lambs [27.88?±?2.85 kg body weight (BW)] were allocated in individual metabolic cages for 20-day adaptation followed by 6 days for measuring dry matter intake (DMI) and CH₄ emission. All lambs received water, mineral supplement, and Cynodon dactylon v. coast-cross hay ad libitum. The treatments consisted of soybean meal (710 g/kg) and ground corn (290 g/kg) [control (CON)]; soybean meal (150 g/kg), ground corn (30 g/kg), and Leucaena hay (820 g/kg) (LEU); soybean meal (160 g/kg), ground corn (150 g/kg), and Mucuna hay (690 g/kg) (STA); and soybean meal (280 g/kg), ground corn (190 g/kg), and Mimosa hay (530 g/kg) (MIM); all calculated to provide 40 g/kg CT (except for CON). DMI (in grams of DMI per kilogram BW per day) was lower for LEU (22.0) than CON (29.3), STA (31.2), and MIM (31.6). The LEU group showed emission of 7.8 g CH₄/day, significantly lower than CON (10.5 g CH₄/day), STA (10.4 g CH₄/day), and MIM (11.3 g CH₄/day). However, when the CH₄ emission per DMI was considered, there were no significant differences among treatments (0.37, 0.36, 0.33, and 0.35 g CH₄/g DMI/kg BW/day, respectively, for CON, LEU, STA, and MIM). The sheep receiving STA had shown a tendency (p?=?0.15) to reduce methane emission when compared to the CON group. Therefore, it is suggested that tropical tanniniferous legumes may have potential to reduce CH₄ emission in sheep, but more research is warranted to confirm these results.     

Genetic immunization with the immunodominant antigen P48 of Mycoplasma agalactiae stimulates a mixed adaptive immune response in BALBc mice

A DNA vaccine against contagious agalactia was developed for the first time, encoding the P48 of Mycoplasma agalactiae. Specific immune responses elicited in BALB/c mice were evaluated. Both total IgG and IgG1 were detected in mice vaccinated with pVAX1/P48. Proliferation of mononuclear cells of the spleen, levels of gamma interferon, interleukin-12, and interleukin-2 mRNAs were enhanced in immunized animals. Results indicate that pVAX1/P48 vaccination induced both T_h1 and T_h2 immune responses. Nucleic acid immunization could be a new strategy against M. agalactiae infections and may be potentially used to develop vaccines for other Mycoplasma diseases.     

Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates

Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa = 0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa = 0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production.     

Patch test reactions to house dust mites in dogs with atopic dermatitis

The purpose of this study was to determine the percentage of dogs with spontaneous atopic dermatitis that show a positive patch test reaction to a commercially available 20% house dust mites mixture containing equal parts of Dermatophagoides farinae and Dermatophagoides pteronyssinus in white petrolatum. In addition, we evaluated whether skin reactions induced after the epicutaneous application of house dust mites were clinically and histologically similar to naturally developed skin lesions of dogs with atopic dermatitis. Furthermore, we investigated if the reactions induced by house dust mites were true allergic reactions by comparing them to atopic lesional skin and to patch test reactions induced by an irritant substance (sodium lauryl sulphate). White petrolatum alone and nonlesional skin sites were used as negative controls. Macroscopic and microscopic evaluations of the patch test and control sites were performed in a blinded fashion at 48 and 72 h after patch test application. Microscopic results were evaluated in a qualitative and quantitative manner. A chi‐square test for homogenicity was used for the quantitative analysis to compare the proportion of each dermal inflammatory cell type among positive histopathological tested sites. P values ≤ 0.05 were considered significant. The study included 12 healthy nonatopic dogs and 13 dogs with nonseasonal atopic dermatitis. None of the nonatopic dogs reacted to house dust mites and white petrolatum. Ten (77%) of the 13 atopic dogs reacted macroscopically and histopathologically to house dust mites. Macroscopic reactions induced by house dust mites were characterized by erythema, oedema and papules. The macroscopic reactions induced by house dust mites were identical to lesional skin in 20% of the dogs and identical to reactions induced by sodium lauryl sulphate in 40% of the dogs. Qualitative histopathological findings showed that the reactions induced by house dust mites were similar to atopic lesional skin in 80% of the dogs and were similar to sodium lauryl sulphate in 20% of the dogs. Quantitative analyses showed that the proportion of neutrophils in reactions induced by sodium lauryl sulphate was significantly higher (P < 0.05) compared to house dust mites reactions, which could be a differentiator factor between an allergic and an irritant reaction. These results showed that the epicutaneous application of house dust mites in dogs with atopic dermatitis induced histopathological lesions similar to spontaneous atopic lesions in dogs. Therefore, this study demonstrated that house dust mites penetrated the skin of dogs with atopic dermatitis and induced an inflammatory response that resembled a true allergic reaction. Funding: Small Companion Animal Grant, University of Minnesota.     

Disseminated pythiosis in three horses

Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures.     

Education and the food-systems veterinarian: the impact of new information technologies

Food systems educators face a double challenge: (1) the inherent change in scope and perspective from raising animals to producing safe food in an environmentally conscientious manner; and (2) the unprecedented demand for higher education, both nationally and internationally. In the modern world, small numbers of producers are capable of feeding a growing population. As demographics have shifted from rural to urban areas, more global livelihoods are derived from manufacturing and services than from agriculture. Education, as one of those services, is accounting for an increasing percentage of world trade, through the physical translocation of students and, more recently, through online education. Within the veterinary realm, colleges outside the United States seek accreditation to better compete for students, and there is increasing pressure from private schools. Today's food systems require a high level of veterinary expertise, with specialization in a particular production system as well as the ability to contribute as part of a larger team that can address economic, bio-security, biological-waste, animal-welfare, food-safety, and public-health concerns. The need for different expertise from food systems specialists (indeed, shortages of all types of veterinary specialists), combined with global competition in education, is a call to action for the veterinary profession. This is an opportunity to revisit and reorganize the delivery of veterinary education, making use of new collaborative technologies for greater efficiency and effectiveness.     

Protein requirements of collared peccary (<Emphasis Type="Italic">Pecari tajacu</Emphasis>)

A nitrogen (N) balance digestion trial was conducted to determine the protein requirement of collared peccaries (Pecari tajacu). In a 4 × 4 Latin square design, four captive adult male peccaries were fed four isoenergy diets containing four different levels of N (11.7, 16.3, 22.8, and 26.7 g N/kg of dry matter—DM). After 15 days of adaptation, a total collection of feces and urine was carried out for five consecutive days. Regression analyses between N intake and N in feces and urine allowed to calculate the metabolic fecal nitrogen (MFN = 2.3 g N/kg of dry matter intake—DMI) and daily endogenous urinary N (EUN = 185 mg N/kg^0.75). Likewise, by regression analyses between consumption of nitrogen and the nitrogen balance (NB = N ingested ? N excreted, mg N/kg^0.75), a daily requirement of 514 mg N/kg^0.75 was calculated. Therefore, if food intake is unrestricted, collared peccaries require a minimum in their diet of about 5.4% crude protein on DM basis. These values are almost as low as those found for browsing and frugivorous wild ruminants, which reinforce the proposition that peccaries’ digestive physiology is nearer to that of domestic and wild ruminants than domestic pigs. This relatively low protein requirement of collared peccary and its great ability to digest protein reveal the relevance of the forestomach for the species on nitrogen/protein metabolism and allow the use of diets with lower crude protein levels than the commercial ones used for the domestic pig, which reduces feed costs.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Seasonal dynamics of the free-living phase of Anocentor nitens at Pedro Leopoldo, Minas Gerais, Brazil

An understanding of seasonal dynamics in the free-living phase of the tropical horse tick, Anocentor nitens, is vital for effective control measures. This study examines seasonal changes in the oviposition and incubation periods, eclosion rates, reproductive success, and larval survival in the free-living phase during the period June 1995-July 1997. Ten engorged female ticks, placed individually in wire-gauze tubes, were exposed in a field of Brachiaria decumbens fortnightly each month, and monitored to measure the duration of the various reproductive parameters, and success. Larval abundance was estimated indirectly, through the percentage of females with viable progeny, and directly by using a flagging method. Duration of the free-living cycle ranged from 12.3 weeks (October and December) to 23.5 weeks (March). The cycle was prolonged in cold and dry months. Larval survival was longer from February to July, and reduced from August to February. Oviposition was recorded in all months, and the egg production index ranged from 26.7 to 47.1%. The eclosion rate ranged from 0.1 to 76.6% in different months. Monthly estimates of larval abundance in the pasture were similar for the two methods used, being higher in months with higher temperatures, relative humidity and rainfall.     

Field evaluation of a coproantigen enzyme-linked immunosorbent assay for diagnosis of canine echinococcosis in a rural Andean village in Peru

One hundred and six dogs (61 males and 45 females) were examined for Echinococcus granulosus infection in a farming cooperative in the central highlands of Peru during November 1998. Canine echinococcosis was diagnosed using direct microscopic examinations of purged feces following arecoline purging and a coproantigen-detection enzyme-linked immunosorbent assay (ELISA) for E. granulosus. Mean age was 2 years with a range of 3 months to 9 years. The overall prevalence of canine echinococcosis using the ELISA test was 79% (84/106). Seventy-four dogs were successfully purged with arecoline. The frequency of canine echinococcosis was 82 (61/74) and 34% (25/74) by the coproantigen ELISA test and arecoline purging, respectively. The sensitivity and specificity of the coproantigen ELISA test was 88 and 95%, respectively. We found this assay to be especially advantageous in remote geographical areas. In future control programs against echinococcosis in Peru and other areas where E. granulosus is endemic the coproantigen ELISA should be used for the surveillance of the dog population.