Kinetics of energy source utilization in Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) embryonic development

The present work evaluates the kinetics of utilization of the main potential energy sources throughout the embryonic developmental stages of Boophilus microplus. The embryonic development of this arthropod is completed in 21 days. Cellularization of the blastoderm occurs on the 6th day and is rapidly followed by germ band extension and segmentation, whose first signs are visible on the 7th day. Cellularization is typically a maternal-driven process, carried out by molecular determinants deposited in the oocyte during oogenesis. On the other hand, segmentation is of zygotic nature, being the consequence of the synthesis of various components by the growing embryo. The enhancement in total B. microplus RNA was observed after cellularization, corroborating the replacement of maternal-driven processes by embryonic zygotic expression. An abrupt increase in oxygen consumption was observed from cellularization until the 8th day of development. The reduction in dry weight at the same period and the susceptibility of oxygen consumption to KCN suggest that the respiration process is activated during early embryonic development. A marked decrease in total lipid content occurred between the 5th and 7th days of development, suggesting this is the main energy source for cellularization. A major reduction in carbohydrate content occurred later, between the 7th and 9th days, and it could be assigned to the morphological segmentation of the embryo. Although the total amount of proteins remains unchanged from oviposition to hatching, a 15% reduction in vitellin (VT) content was observed before cellularization, up to the 4th day after egglaying. This observation was correlated to the synthesis of new proteins needed to support early embryo development. Additional 20% of VT was consumed thereafter, mainly at the end of embryogenesis, and in this case VT is probably used as energy source to the older embryo. Altogether, these data indicate different energy sources for maternal and zygotic driven processes.     

Diurnal and nocturnal herbivore induction on maize elicit different innate response of the fall armyworm parasitoid, <Emphasis Type="Italic">Campoletis flavicincta</Emphasis>

Herbivore-attacked plants produce specific volatile substances that represent important cues for host finding by natural enemies. The fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is a voracious herbivore and usually feed on maize in all periods of the day. Given that plant needs light to synthesize de novo herbivore-induced volatiles, volatile blend may be changed depending on time of the day the plant is induced, what could interfere in natural enemy foraging. In this sense, the current study aimed to investigate differential attractiveness of maize elicited by fall armyworm regurgitant under light and dark conditions to its specialist larval parasitoid Campoletis flavicincta (Ashmead) (Hymenoptera: Ichneumonidae). All bioassays were conducted in Y-tube olfactometer to assess parasitoid response to odors from undamaged maize, mechanical damage, and regurgitant-treated plants at 0–1, 5–6, and 24–25 h after induction. The results showed that naïve wasps were attracted to volatiles emitted by nocturnal regurgitant-treated maize at 5–6 h, but not to odors from diurnal regurgitant-treated plants. The differential attractiveness is likely due to blend composition as nocturnal regurgitant-treated plants emit aromatic compounds and the homoterpene (3E)-4,8-dimethyl-1,3,7-nonatriene in larger amounts than diurnal-treated plants.     

Platelet-Rich Plasma in the Treatment of Induced Tendinopathy in Horses: Histologic Evaluation

This study was carried out to evaluate the effect of platelet-rich plasma (PRP) on the treatment of tendinopathy induced in the superficial digital flexor tendon (TFDS) of horses, by using histologic evaluation. Six healthy crossbred geldings aged 8 to 15 years (12 ± 3) were used. The TFDS tendinopathy was provoked in both forelimbs, by intratendinous administration of 2.5 mg collagenase (2.5 mg/mL), and this procedure was considered as the beginning of the experimental phase. At 12 days after induction of the tendinopathy, the animals were subjected to the following treatments: (1) in the lesion caused in the right superficial digital flexor tendon (PRP-treated group), 2.5 mL PRP activated with calcium chloride at 0.0125 mol/L at concentrations from 320,000 to 500,000 platelets/μL, were injected; (2) in the tendinopathy of the left SDFT (control group), 2.5 mL 0.9% saline solution was administrated. Thirty-six days after the treatments, a biopsy of the injured area was performed for histologic evaluation. In both groups, the histologic analysis showed an increase in the fibroblastic density, as well as the presence of neovascularization, lymphocytes, and plasmocytes infiltrate and tissue organization at variable intensity. In the PRP-treated group, the SDFT was more organized, with the collagen fibers and fibroblasts being better arranged on the tendon matrix. The numbers of the fibroblasts and blood vessels did not differ between the groups. Histologic evaluation 36 days after tendinopathy showed that injuries under a single PRP treatment present a more uniform and organized tissue repair when compared with the control group.     

Shortening the Postpartum Anoestrous Interval in Suckled Crossbred Dual Purpose Cows Using Progestagen Intravaginal Sponges plus eCG and PGF2α

One hundred and twenty‐six suckled crossbred cows (Bos taurus × Bos indicus), with body condition score ≥3 (1–5 point scale), were employed in the present study to evaluate the effectiveness of intravaginal progestin‐releasing sponges (IVS) for shortening anoestrous interval. Fifty‐four cows were assigned to control group. Seventy‐two cows were treated with IVS impregnated with 250 mg of medroxy‐acetate‐progesterone (MAP) as follows: day 0, IVS plus 5 mg of 17β‐E and 50 mg of MAP i.m.; day 6, 500 IU of equine chorionic gonadotropin and 25 mg prostaglandin F_2α i.m.; day 8, IVS withdrawal and day 9, 1 mg 17β‐E i.m. Cows were also grouped according to postpartum days (dpp) at treatment: MAP <70 days (n = 25); control <70 days (n = 22); MAP >70 days (n = 47); control >70 days (n = 32). From IVS removal, cows were detected in oestrus and inseminated. Cows not detected in oestrus were timed artificial insemination 72 h after sponge removal. Treatment effect on oestrous rate (ER), conception rate (CR), pregnancy rate (PR) and treatment to conception intervals (TCI) and calving to conception intervals (CCI) were evaluated. The ER, CR and PR were analysed using proc logistic , while TCI and CCI with proc glm of SAS. The groups MAP <70 days and MAP >70 days showed higher (p < 0.01) ER than control <70 days and control >70 days (84.0% and 76.6% vs 31.8% and 31.3% respectively). The PR was higher (p < 0.01) in MAP <70 days vs control <70 days (64.0% vs 22.7%) and also higher (p < 0.05) in MAP >70 days vs control <70 days (40.4% vs 18.8%). The TCI and CCI were shorter (p < 0.01) in MAP <70 days vs control <70 days (36.0 and 95.8 days; 95.3 and 158.6 days respectively). In conclusion, only cows treated with IVS before 70 dpp had a CCI shorter than 100 days, consequently this treatment shortened postpartum anoestrous interval in crossbred dual purpose cattle.     

Quantifying Microbial Methane Oxidation Efficiencies in Two Experimental Landfill Biocovers Using Stable Isotopes

Stable isotope analyses were performed on gas samples collected within two instrumented biocovers, with the goal of evaluating CH₄ oxidation efficiencies (f ₀). In each of the biocovers, gas probes were installed at four locations and at several depths. One of the biocovers was fed with biogas directly from the waste mass, whereas the other was fed through a gas distribution system that allowed monitoring of biogas fluxes. While the f ₀ values obtained at a depth of 0.1 m were low (between 0.0% and 25.2%) for profiles with poor aeration, they were high for profiles with better aeration, reaching 89.7%. Several interrelated factors affecting aeration seem to be influencing f ₀, including the degree of water saturation, the magnitude of the biogas flux and the temperature within the substrate. Low f ₀ values do not mean necessarily that little CH₄ was oxidized. In fact, in certain cases where the CH₄ loading was high, the absolute amount of CH₄ oxidized was quite high and comparable to the rate of CH₄ oxidation for cases with low CH₄ loading and high f ₀. For the experimental biocover for which the CH₄ loading was known, the oxidation efficiency obtained using stable isotopes (f ₀?=?55.67% for samples taken inside flux chambers) was compared to the value obtained by mass balance (f ₀?=?70.0%). Several factors can explain this discrepancy, including the high sensitivity of f ₀ to slight changes in the isotopic fractionation factor for bacterial oxidation, α _ox, uncertainties related to mass flow metre readings and to the static chamber method.     

Effect of fertilizers, lime, and inoculation with rhizobia and mycorrhizal fungi on the growth of four leguminous tree species in a low-fertility soil

The aim of this study was to evaluate the effects of lime, fertilizers, mycorrhizal fungi, and selected rhizobia strains on the growth of four woody legume species, Albizia lebbeck (L.) Benth., Enterolobium contortisiliquum (Vell.) Morong., Leucaena leucocephala (Lam.) de Wit, and Sesbania virgata (Cav.) Pers. in a low-fertility soil. The experiment was conducted under greenhouse condition in plastic pots (4 kg). Eight treatments and eight replicates per treatment were performed in a completely randomized design. The treatments were: (1) complete treatment (C) (NPK fertilization?+?micronutrients?+?liming?+?MR that is inoculation with mycorrhizal fungi and rhizobia); (2) C minus N (C???N that is as C without the addition of N); (3) C???N???M (as C???N without inoculation with arbuscular mycorrhizal fungi (AMF)); (4) C???N??R (as C???N without inoculation of rhizobia); (5) C???N???liming (as C???N without liming); (6) C???N???micro (as C???N without addition of micronutrients); (7) C???N???P (as C???N without addition of P); (8) control without fertilization, liming, and without inoculation with AMF and rhizobia. After 4 months of growth, we determined the yield of individual plants, nodulation, mycorrhizal colonization, and nutrient contents. Phosphorus was the most limiting nutrient for plant growth, followed by nitrogen. L. leucocephala and S. virgata had the most robust response to the addition of micronutrients and liming, showing an increase in nutrient content, plant height, and root and shoot dry matter. When compared to the single inoculation, the dual inoculation increased growth of all plants, except that of A. lebbeck, which did not respond to either rhizobia or mycorrhizal fungi inoculation.     

Molecular typing of Mycobacterium bovis isolates from south-east Brazil by spoligotyping and RFLP

The identification of 163 strains of Mycobacterium bovis by polymerase chain reaction (PCR) and microbiological tests was carried out on 252 tuberculous-like lesions (TLLs) collected from slaughtered cattle in south-east Brazil. This study compared the usefulness of three genotyping techniques, IS6110-restriction fragment length polymorphism (RFLP), polymorphic guanine-cytosine-rich sequence (PGRS)-RFLP and direct repeat (DR)-spoligotyping, as applied to M. bovis isolates. Based on IS6110-RFLP genotyping we selected a group of 23 isolates containing more than one IS6110 copy, along with 16 samples containing one IS6110 copy from different geographical areas, evenly distributed among dairy (eight) and beef cattle (eight). These selected isolates were analysed by PGRS-RFLP and DR-spoligotyping genotyping. Dairy cattle (17%) display a higher frequency of multiple IS6110 copies than beef cattle (10%). A comparison between the genotype data obtained fails to show a correlation between the main clusters found by the three techniques. However, the clustering of each genotyping procedure revealed that the majority of strains are closely related. The RFLP-PGRS patterns showed a sizable group (20.5%) containing a 5.5 kb fragment and the predominant spoligotype is similar to that from the BCG vaccine strain. Unexpectedly, four strains (2.4%) showed drug resistance to 0.2 microg/ml isoniazid and 20 microg/ml ethionamide, but none of them was resistant to rifampicin or other antibiotics tested.     

Molecular Typing of Mycobacterium bovis Isolates from South‐east Brazil by Spoligotyping and RFLP

The identification of 163 strains of Mycobacterium bovis by polymerase chain reaction (PCR) and microbiological tests was carried out on 252 tuberculous‐like lesions (TLLs) collected from slaughtered cattle in south‐east Brazil. This study compared the usefulness of three genotyping techniques, IS6110‐restriction fragment length polymorphism (RFLP), polymorphic guanine‐cytosine‐rich sequence (PGRS)‐RFLP and direct repeat (DR)‐spoligotyping, as applied to M. bovis isolates. Based on IS6110‐RFLP genotyping we selected a group of 23 isolates containing more than one IS6110 copy, along with 16 samples containing one IS6110 copy from different geographical areas, evenly distributed among dairy (eight) and beef cattle (eight). These selected isolates were analysed by PGRS‐RFLP and DR‐spoligotyping genotyping. Dairy cattle (17%) display a higher frequency of multiple IS6110 copies than beef cattle (10%). A comparison between the genotype data obtained fails to show a correlation between the main clusters found by the three techniques. However, the clustering of each genotyping procedure revealed that the majority of strains are closely related. The RFLP‐PGRS patterns showed a sizable group (20.5%) containing a 5.5 kb fragment and the predominant spoligotype is similar to that from the BCG vaccine strain. Unexpectedly, four strains (2.4%) showed drug resistance to 0.2 μg/ml isoniazid and 20 μg/ml ethionamide, but none of them was resistant to rifampicin or other antibiotics tested.