Effect of oil-based formulations of acaripathogenic fungi to control Rhipicephalus microplus ticks under laboratory conditions

The formulations of acaripathogenic fungi to control ticks have been widely studied. The present study evaluated the efficacy of oil-based formulations of Metarhizium anisopliae sensu lato (s.l.), isolate Ma 959, and Beauveria bassiana, isolate Bb 986, on different Rhipicephalus microplus stages, comparing the efficacy between aqueous suspensions and 10, 15 and 20% mineral oil formulations. Twelve groups were formed: one aqueous control group; three mineral oil control groups, at 10, 15 or 20%; two aqueous fungal suspensions of M. anisopliae s.l. or B. bassiana; and three formulations of M. anisopliae (s.l.) or B. bassiana containing 10, 15, and 20% mineral oil. To prepare aqueous suspensions and oily formulations, fungal isolates were cultivated on rice grains in polypropylene bags. The conidial suspensions and formulations had a concentration of 10(8)conidia/mL. Bioassays were repeated twice. After treatment, the following biological parameters of engorged females were evaluated: hatching percentage, egg production index, nutritional index, and percentage of tick control. The following parameters were evaluated in the bioassays with eggs: period of incubation, period of hatch, and hatching percentage. Mortality was evaluated in bioassays with larvae. M. anisopliae s.l. and B. bassiana oil-based formulations were more effective than aqueous suspensions against R. microplus eggs, larvae and engorged females, however, there was no significant difference between the three oil concentrations used. M. anisopliae s.l. and B. bassiana formulated in mineral oil reached 93.69% and 21.67% efficacy, respectively, while M. anisopliae s.l. and B. bassiana aqueous suspensions attained 18.70% and 1.72% efficacy, respectively. M. anisopliae s.l. oil-based formulations caused significant effects in all biological parameters of engorged females while B. bassiana oil-based formulations modified significantly the nutritional index only. Eggs treated with M. anisopliae s.l. and B. bassiana oil-based formulations showed hatching rates that decreased 102.5 and 3.65 times, respectively. In the bioassay with larvae, M. anisopliae s.l. oil-based formulations caused nearly 100% mortality five days after treatment, while larva treated with B. bassiana oil-based formulations reached 100% mortality at day 20 after treatment. Larva from oil-based control groups showed mortality at day 15 after treatment, which indicated a possible toxic effect of the oil for this R. microplus stage. The results showed that the fungal mineral oil formulations tested were more effective than the aqueous suspension. Oil-based formulations at 10, 15 and 20% enhanced the activity of M. anisopliae s.l. Ma 959, and B. bassiana Bb 986, isolates against R. microplus eggs, larvae, and engorged females tick. Mineral oil was effective as an adjuvant in formulations of M. anisopliae s.l., Ma 959, and B. bassiana, Bb 986, for the control of R. microplus under laboratory conditions.     

Influences of Sex and Age on the Hematological Profile of the Jundiá (Silver Catfish) Rhamdia quelen

In this study, sex and age influenced the hematological profiles of Jundiá (Silver Catfish) Rhamdia quelen. Females showed lower levels of hemoglobin, young fish increased lymphocyte counts, and older fish increased hematocrit values. These results indicate that, depending on age and sex, the fish have disparate hematological profiles. For this reason, it is important to consider the sex and age of an R. quelen when examining the impact of environmental and management factors on this species in terms of their hematological profiles.

Received May 24, 2015; accepted March 24, 2016 Published online August 2, 2016     

Contrasting effects of heat shock during in vitro maturation on development of in vitro‐fertilized and parthenogenetic bovine embryos

This study investigated the influence of heat shock during in vitro maturation on embryo development following in vitro fertilization (IVF) or parthenogenesis (Part). Immature bovine cumulus–oocyte complexes were exposed to heat shock (41.0°C) during the first 12 hr of in vitro maturation (IVM), followed by 12 hr at 38.5°C. Control group consisted of in vitro maturation for 24 hr at 38.5°C. Oocytes were in vitro‐fertilized or activated with ionomycin and cultured in vitro for 192 hr post‐in vitro insemination or parthenogenetic activation (hpia). There was an interaction (p < .01) between temperature of IVM and method of oocyte activation (IVF or Part) for cleavage at 48 hpia. Heat shock had a negative impact (p < .01) on cleavage of IVF embryos, whereas no (p > .05) effect was found in the Part embryos. Embryo development towards blastocyst stage at 168 and 192 hpia decreased in both IVF and Part embryos derived from heat‐shocked oocytes. Heat shock increased (p < .05) the apoptotic index in Part blastocysts, but no effect (p > .05) was found in IVF counterparts. Heat shock also down‐regulated the expression of AQP3 (p < .01) and up‐regulated the expression of HSP70.1 (p < .01) in Part blastocysts, whereas it down‐regulated the expression of ATP1A1 (p < .05) in IVF blastocysts. In conclusion, the effects of heat shock during IVM on early embryo cleavage and blastocyst apoptosis are influenced by the method of oocyte activation and expression of some genes can be disturbed in embryos derived from heat‐shocked oocytes.     

Mapping genomic regions for reproductive traits in beef cattle: Inclusion of the X chromosome

Although the second largest chromosome of the genome, the X chromosome is usually excluded from genome-wide association studies (GWAS). Considering the presence and importance of genes on this chromosome that are involved in reproduction, the aim of this study was to evaluate the effect of its inclusion in GWAS on reproductive traits (scrotal circumference [SC], early pregnancy [P16] and age at first calving [AFC]) in a Nelore herd. Genotype data from 3,263 animals with the above-mentioned phenotypes were used. The results showed an increase in the variances explained by the autosomal markers for all traits when the X chromosome was not included. For SC, there was an increase of more than 10% for the windows on chromosomes 2 and 6. For P16, the effect was increased by almost 20% for windows on chromosome 5. The same pattern was found for AFC, with an increase of more than 10% for the most important windows. The results indicate that the noninclusion of the X chromosome can overestimate the effects of autosomes on SC, P16 and AFC not only because of the additive effect of the X chromosome itself but also because of its epistatic effect on autosomal genes.     

SHORT COMMUNICATION: Neutrophil oxidative metabolism and haemogram of sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E

The objective of this study was to evaluate neutrophil oxidative metabolism and haemogram in sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E. Twenty male Corriedale sheep were utilised and distributed into four experimental groups each with five animals infected with larvae: G1‐ supplemented with sodium selenite, 0.2 mg/kg body weight (bw) given intramuscularly (IM); G2‐ supplemented with sodium selenite and vitamin E, 0.2 mg/kg bw and 2000 IU per animal, respectively, both IM; G3‐ supplemented with vitamin E, 2000 IU per animal IM; G4‐ not supplemented. A haemogram and the number of parasite eggs were determined in samples of blood and faeces, respectively, on days zero (T0), 20 (T1), 40 (T2) and 60 (T3) and nitroblue tetrazolium (NBT) assays were performed on heparinized blood samples taken on days zero, 30 and 60. A lower total leucocyte count was detected in G1 in relation to G4 at T4. Lymphocytes were reduced in G1 in relation to G3 and G4 at T3. In both non‐stimulated (NBT‐NS) and stimulated (NBT‐S) dye reduction assays, there was reduced activity at 60 days, in relation to other times in the groups treated with selenium (G1 and G2). On the basis of results obtained, we conclude that supplementation with selenium provides better antioxidant protection to neutrophils.     

Early detection of Neophysopella tropicalis in grapevine leaves and on spore traps by qPCR

Neophysopella tropicalis, one of the causal agents of Asian grapevine leaf rust (AGLR), can cause severe epidemics in Brazil that lead to yield losses in commercial vineyards. An early detection of the pathogen by air sampling of urediniospores on spore traps or in symptomless leaves would be valuable to multiple studies, such as epidemics modelling, risk forecasting, monitoring of pathogen introductions in rust-free areas, and predicting the beginning of epidemics. This study developed a quantitative PCR (qPCR) protocol to quantify N. tropicalis urediniospores attached to adhesive tapes and in grapevine leaves before symptom appearance. A specific primer pair was designed based on the internal transcribed spacer (ITS) sequence region of the AGLR pathogen. Standard amplification curves using genomic DNA from urediniospores of N. tropicalis and from urediniospores attached to adhesive tapes were established. Grapevine leaves inoculated with N. tropicalis were collected at 2, 5, and 7 days postinoculation (dpi). One primer pair (580F/720R) amplified a 140 bp product in all AGLR isolates but did not amplify products of other rust genera, such as Phakopsora, Puccinia, Hemileia, Tranzschelia, Cerotelium, and Coleosporium. As little as 0.1 pg DNA and 10 urediniospores of N. tropicalis attached to adhesive tapes could be detected. qPCR enabled the detection of the pathogen as early as 2 dpi, before symptom appearance. This method can be used to monitor N. tropicalis inoculum in grapevine-growing areas and to quantify symptomless infections of the AGLR pathogen.     

Influence of phosphorus fertilization on the response of pinus genotypes to glyphosate subdoses

New Forests - Previous studies have reported that phosphorus transporters are involved in glyphosate uptake by plants, so phosphorus fertilization might act as an attenuator of glyphosate effects...     

Detection, quantification, and pharmacokinetics of furosemide and its effects on urinary specific gravity following IV administration to horses

Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.     

Pine branch of Araucaria angustifolia in horse bronchi: A series of four cases

Horses at pasture are susceptible to the aspiration of foreign bodies. In Curitiba, southern Brazil, four horses of different breeds and ages were examined with regards to a persistent cough that lasted for several months, fetid breath, and no fever. The animals were referred for bronchoscopic examination performed under sedation and tracheobronchial topical anaesthesia. Bronchoscopies in all four horses revealed a pine branch of the species Araucaria angustifolia, native to southern Brazil, in the lumen of a third to fifth generation bronchus. Complete removal of the branch was possible in one case using a polypectomy loop inserted through the biopsy channel of the bronchoscope. In two cases, partial removal was possible. In one case the branch was adhered to the swollen mucosa and could not be extracted; this animal is being kept at pasture with intermittent antibiotic treatment as necessary. In conclusion, foreign bodies consisting of pine branches in the tracheobronchial tree of horses may be more common than currently recognised, as Araucaria genus is present in other countries and should be suspected in horses with chronic coughing associated with fetid breath and a history of reoccurrence following symptomatic treatments.