Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Search for bovine papilloma virus DNA in bovine ocular squamous cell carcinomas (BOSCC) and BOSCC-derived cell lines.

Although the cause of bovine ocular squamous cell carcinoma (BOSCC) is attributed to viruses in addition to cofactors (eg, UV light), to our knowledge, the final causative agent has not been described. Bovine papilloma virus (BPV)-like particles were detected in approximately 33% of various putative precursor lesions of BOSCC. In contrast, it was reported that, using BPV-specific antibodies, it was not possible to detect viral antigens in BOSCC. Fourteen established BOSCC and 9 BOSCC-derived cell lines were examined for BPV DNA. Probes of all 6 known BPV types were used in various hybridization assays. Neither Southern blot analysis, under high and low stringency conditions, nor in situ hybridization resulted in detection of BPV DNA. Papilloma viruses were not observed in electron microscopic studies. Results exclude direct association between BOSCC and BPV types 1 to 6, or as yet unknown closely related BPV types. However, BPV may contribute to induction of precursor lesions or events leading to carcinogenic transformation, without being relevant for maintenance of the tumor.     

Pseudorabies virus infections in explants of porcine nasal mucosa.

The spread of infection and the morphogenesis of three pseudorabies virus strains were studied in explants of porcine nasal mucosa. Virulent NIA-3 virus was compared with a deletion mutant 2.4N3A, and with a non-virulent Bartha virus. All three virus strains infected nasal epithelial cells. NIA-3 virus particles were enveloped mainly at the inner nuclear membrane; the virus rapidly invaded the stroma, causing widespread necrosis. In contrast, 2.4N3A virus particles were enveloped mainly at the endoplasmic reticulum and the infection spread more slowly. Bartha virus particles were enveloped mainly at the endoplasmic reticulum; the infection spread slowly and remained restricted to the epithelial cells. In situ DNA hybridisation showed an accumulation of Bartha virus DNA in the nucleus 24 hours after inoculation. In nasal mucosa viral virulence seemed directly related to the speed of replication and release of virus from infected cells.     

Effects of Melatonin Implants During Non‐Breeding Season on Sperm Motility and Reproductive Parameters in Rasa Aragonesa Rams

The effect of melatonin implants administered during non‐breeding season in Rasa Aragonesa rams on sperm motility parameters and other reproductive traits was assessed. In a first experiment, two Rasa Aragonesa rams were implanted (with melatonin group M), remaining other two males as control group (C). Semen of each group was collected from 1 May to 23 June, twice or three times a week, and motility parameters were assessed using a computer‐assisted sperm analysis system. Melatonin increased the percentage of progressive motile spermatozoa, particularly during 46–75 days after melatonin implantation (p < 0.01). In experiment 2, M and C in vitro fertilization ability had been determined by zona‐pellucida binding assays, using spermatozoa from experiment 1, obtained 60–70 days after melatonin was implanted. A significantly higher number of spermatozoa attached per oocyte was observed in frozen‐thawed immature ovine oocytes incubated with sperm from M animals than in those incubated with sperm from the C group (p < 0.01). Finally, a field assay (experiment 3) was performed. In this case, five Rasa Aragonesa rams were implanted with melatonin and three remained as control group. Sperm doses from those animals were used for artificial insemination of 2608 Rasa Aragonesa ewes from 39 different farms at non‐breeding season. Fertility, litter size and fecundity were studied. Semen from melatonin implanted rams seemed to increase both fertility and fecundity in ewes inseminated with spermatozoa obtained 46–60 days after implantation (p < 0.1). Thus, melatonin treatment in rams during non‐breeding season modifies sperm motility parameters and seems to improve the fertilization parameters obtained.     

Cowdria ruminantium is recognized by a monoclonal antibody directed against the major outer membrane protein of Chlamydia trachomatis.

The relationship between Cowdria ruminantium and Chlamydia trachomatis was studied by immunofluorescence. A monoclonal antibody directed against the major outer membrane protein of C. trachomatis recognized rickettsial colonies of C. ruminantium in infected goat brain. No specific fluorescence was observed in non-infected brain. Two commercial Chlamydia-specific monoclonal antibodies as well as polyvalent anti-Chlamydia rabbit serum recognized C. trachomatis, but did not recognize Cowdria. Moreover, polyvalent Cowdria antiserum failed to recognize C. trachomatis cultivated in HeLa cells. It is concluded that Cowdria and Chlamydia are to a certain extent related, confirming similarities in ultrastructure and developmental cycle.     

Efficacy of adding chemical and microbial additives to silage on beef cattle performance: Systematic review and meta-analysis

The effects of different additives on farm-scale silage quality and beef cattle performance are inconsistent. This study aimed to carry out a systematic review and meta-analysis to evaluate the efficacy of chemical and microbial additives to silage on beef cattle performance. Systematic searches were performed using databases and scientific journals, and 42 articles were selected. Data for all variables were grouped into subgroups according to the additive type. For dry matter intake and average daily gain, the data were also grouped by forage type due to greater comparison numbers. The treatment mean differences and 95% confidence intervals (p < 0.05) were analysed using a random-effects model. The use of homo- and heterofermentative microbial inoculant mixtures and chemical additives (Ch) increased the average daily gain of beef cattle fed maize/sorghum silage. Homofermentative microbial inoculant (Ho), Ch, and a mixture of microbial inoculant and chemical additives also increased the average daily gain of beef cattle fed temperate grasses. Only Ch increased dry matter intake. Ch increased feed efficiency, and Ch and Ho increased carcass weight. The evaluated additives improved the silage fermentation process mainly via pH and ammonia nitrogen reduction. Overall, this meta-analysis demonstrated that silage additives improved the ensiling process and beef cattle performance, with better results with Ch use. Due to the aerobic stability and microbiological profile analyses being carried out more in laboratory-scale silos, more studies are needed to determine these silage parameters after opening the silo at the farm scale.     

Influence of native or exotic trees on soil fertility in decades of silvopastoral system at the Brazilian savannah biome

Silvopastoral systems (SPSs) are considered a conservationist system by combining forestry species cultivation with domesticated grazing animals, intercropping trees, and pasture in the same area. SPSs are noun for its contribution to improve soil chemical and physical properties, enhance grass nutritional status, provide healthier animal environment, and increase farmers’ options to maximize the use of land. In soil degraded areas, such as those found in Brazilian savannah biome (BSB), the SPSs constitute an alternative to soil recuperation in these areas. Therefore, our aim was to assess soil fertility attributes at different depths on a BSB grown with Brachiaria brizantha cv. Marandu grass, the influence of SPS with native tree zeyheria (Zeyheria tuberculosa) settled in 1984, or with the exotic eucalyptus (Eucalyptus grandis) settled in 1994. The soil water pH, P, Ca, Mg, and K contents, as well as soil base sum and saturation, were found superior mostly at the top layer (0–2 cm) of the eucalyptus site, than for the ipê-felpudo site. This response was correlated to greater soil organic matter and carbon at the eucalyptus site, which demonstrated to have high nutrient cycling rates. Around 50 % of the CEC in both SPSs was occupied by bases, demonstrating that the areas have large potential to absorb applied nutrients. The SPSs have no significant impact on S-SO₄ ^2? or micronutrient availability. The litter composition in eucalyptus site was accountable for the superior results of this SPS, which has shown as a promising choice for land use and conservation in the BSB.     

Standard area diagram set to assess rust severity on eucalyptus leaves

This study aimed to develop and validate a standard area diagram set (SADs) to help assess rust severity caused by Austropuccinia psidii on eucalyptus leaves. The SADs has eight colour images of leaves with severity values ranging from 0.97% to 37.3%. The SADs were validated by 16 raters, who assessed the same set of 50 images twice: the first without the SADs and the second using it as an aid. Accuracy and precision of the estimates were determined based on Lin's concordance correlation coefficient, and the inter‐rater reliability was measured using the coefficient of determination (R²) and intraclass correlation. The estimates of severity of eucalyptus rust were more accurate, precise and reliable using SADs. These results demonstrate the value of SADs for several activities, including epidemiological studies, treatments comparison, disease monitoring, decision‐making in eucalyptus rust management measures, and in breeding programs.