Microbiological quality of Litopenaeus vannamei culture using conventional and biofloc systems

Shrimp farming is a fast‐expanding activity that has supported the growth in the supply of these crustaceans to consumers around the world. However, the end product is vulnerable to contamination at all stages of the process, including the rearing tanks, where current practices prioritize to raise stocking densities and the minimization of water renewal. It is thus important to evaluate the potential of these systems for the proliferation of undesirable microorganisms, which may render the product unfit for human consumption. In the present study, the presence of coagulase‐positive Staphylococcus, Salmonella spp., Vibrio spp., and total and thermotolerant coliforms was verified in biofloc tank and conventional pond systems used for the rearing of Litopenaeus vannamei in Pernambuco, Brazil, and the results were compared with the legislation regulating the marketing of fresh shrimp. Samples were collected from two biofloc tanks with a density of 375 shrimp m^?2, and two conventional ponds with 12 shrimp m^?2. None of the samples tested positive for either Salmonella spp. or coagulase‐positive Staphylococcus, which is consistent with the legislation. While no standards are defined legally for Vibrio spp. or coliforms, very low concentrations were recorded in both systems, in comparison with other studies. While some variation in bacterial contamination was observed over the rearing process, the end product of both systems was fit for human consumption. The results of the study indicate that, while the water is not renewed in the biofloc system, the development of undesirable microorganisms can be controlled, with no adverse effects for the end product.     

Selection of cassava accessions with multiple resistance to pathogens associated with root rot disease

Cassava root rot disease (CRRD) severely affects productivity in several countries. The objective of this study was to estimate genetic parameters and to identify multiple resistance sources against pathogens associated with CRRD. The symptoms caused by Fusarium spp., Phytophthora spp., and Botryosphaeriaceae species in peel and pulp from the roots were evaluated in 277 accessions using a whole tuberous root inoculation method. The resistance data were obtained by REML/BLUP (restricted maximum likelihood/best linear unbiased predictor). The classic selection index (CI), multiplicative (MI), and sum of ranks (SRI) were used to identify the accessions with multiple resistance. For all pathogens, the environmental variance (\(\sigma_{e}^{2}\)) was the most important component. Individual heritability \(\left( {h_{g}^{2} } \right)\) was of low magnitude for resistance to most pathogens (0.16 ± 0.02—peel and 0.31 ± 0.03—pulp for Fusarium spp.; 0.26 ± 0.03—peel and 0.30 ± 0.03—pulp for Phytophthora spp.; and 0.28 ± 0.03—peel and 0.27 ± 0.03—pulp for Botryosphaereacea species). The distribution of CRRD symptoms indicated the presence of quantitative inheritance. The direct selection of the 15 more resistant accessions based on the genotypic predicted values result in high reductions of disease (>50%). However, there was a low matching rate of the most resistant accessions for each pathogen and the different parts of the tuberous roots (peel and pulp). The CI and MI were the most promising compared to the SRI to ensure high and balanced resistance for each pathogen. Understanding the genetic basis of resistance to CRRD and the identification of sources with multiple resistance may be useful in various management strategies to control the disease.     

Genetic variation of germination cold tolerance in Japanese rice germplasm

Low temperatures at the initial stages of rice development prevent fast germination and seedling establishment and may cause significant productivity losses. In order to develop rice cultivars exhibiting cold tolerance, it is necessary to investigate genetic resources, providing basic knowledge to allow the introduction of genes involved in low temperature germination ability from accessions into elite cultivars. Japanese rice accessions were evaluated at the germination under two conditions: 13°C for 28 days (cold stress) and 28°C for seven days (optimal temperature). The traits studied were coleoptile and radicle length under optimal temperature, coleoptile and radicle length under cold and percentage of the reduction in coleptile and radicle length due to low temperature. Among the accessions studied, genetic variation for traits related to germination under low temperatures was observed and accessions exhibiting adequate performance for all investigated traits were identified. The use of multivariate analysis allowed the identification of the genotypes displaying cold tolerance by smaller reductions in coleoptile and radicle lenght in the presence of cold and high vigour, by higher coleoptile and radicle growth under cold.     

Natural co-infection of torque teno virus and porcine circovirus 2 in the reproductive apparatus of swine

This work aimed to detect and study natural co-infection of Circoviridae torque teno virus (TTV) and porcine circovirus 2 (PCV2) in the swine reproductive apparatus. Semen and organs from 17 boars were tested by nested and real-time PCR. PCV2 was amplified from semen (47%), lymph nodes (84.6%) and testicles (35.3%). TTV2 was amplified from 16/17 testis and 13/13 lymph nodes. TTV1 DNA was detected in fewer testicle samples (2/17), which were also TTV2 positive. Analyzed ovaries, follicular fluid and uteri of 83 culled sows showed TTV2, TTV1 and PCV2 from 49.3%, 30.1% and 6.0% of the sows, respectively. Sperm analysis indicated insignificant differences between PCV2 and TTVs positive and negative boars. The most frequent pathologic lesion in sows was endometritis (28.9%), but this was unassociated with PCV2 or TTVs detection. These findings question the importance of PCV2 and TTV2 natural co-infection in the pathology of porcine reproductive failures.     

Leishmanicidal activity in vitro of Musa paradisiaca L. and Spondias mombin L. fractions

Visceral leishmaniasis (VL) is a zoonotic disease characterized by infection of mononuclear phagocytes by Leishmania chagasi. The primary vector is Lutzomyia longipalpis and the dog is the main domestic reservoir. The control and current treatment of dogs using synthetic drugs have not shown effectiveness in reducing the incidence of disease in man. In attempt to find new compounds with leishmanicidal action, plant secondary metabolites have been studied in search of treatments of VL. This study aimed to evaluate the leishmanicidal activity of Musa paradisiaca (banana tree) and Spondias mombin (cajazeira) chemical constituents on promastigotes and amastigotes of L. chagasi. Phytochemical analysis by column chromatography was performed on ethanol extracts of two plants and fractions were isolated. Thin layer chromatography was used to compare the fractions and for isolation the substances to be used in vitro tests. The in vitro tests on promastigotes of L. chagasi used the MTT colorimetric method and the method of ELISA in situ was used against amastigotes besides the cytotoxicity in RAW 264.7 cells. Of the eight fractions tested, Sm1 and Sm2 from S. mombin had no action against promastigotes, but had good activity against amastigotes. The fractions Mp1 e Mp4 of M. paradisiaca were very cytotoxic to RAW 264.7 cells. The best result was obtained with the fraction Sm3 from S. mombin with IC(50) of 11.26 μg/ml against promastigotes and amastigotes of 0.27 μg/ml. The fraction Sm3 characterized as tannic acid showed the best results against both forms of Leishmania being a good candidate for evaluation in in vivo tests.     

Acetylcholinesterase activity and lipid peroxidation in the brain and spinal cord of rats infected with Trypanosoma evansi

Neurological and locomotor clinical signs are described in animals infected with Trypanosoma evansi. These disturbances may be related to changes in the amount of acetylcholine (neurotransmitter) in the synaptic cleft. Therefore, changes in acetylcholinesterase (AChE) activity and lipid peroxidation in brain and spinal cord of T. evansi-infected rats were investigated. Each rat was intraperitoneally infected with 10(6) trypomastigotes kept in fresh (group A; n=13) and cryopreserved blood (group B; n=13). Thirteen served as uninfected (not-infected; group C). In days 4 and 30 post-infection (PI) the rats were anesthetized and subsequently decapitated to obtain the brain and the spinal cord (between vertebrae L1 and S2). The brain was removed and dissected (cerebellum, cerebral cortex, striatum and hippocampus) to measure the activity of AChE and lipid peroxidation, determined by TBARS levels. To verify if T. evansi was present in the central nervous system (CNS), brain structures of three rats of each group were processed by PCR T. evansi-specific. AChE activity was significantly increased in all brain structures and decrease in spinal cord in infected rats in 4 PI (P<0.05). The levels of TBARS were decreased in the brain structures, differently from spinal cord, which showed increased lipid peroxidation in 4 PI. The AChE activity in striatum, cerebral cortex, hippocampus and spinal cord reduced concomitantly with the increase of the enzyme in cerebellum of the infected rats (P<0.05), and the TBARS levels increased in cerebellum, striatum and spinal cord of infected rats compared to non-infected animals in 30 PI. The PCR was positive for T. evansi in all structures of the brain, confirming the presence of the parasite in the CNS. Based on the results, we conclude that the changes in AChE activity and lipid peroxidation in the CNS are induced by infection with T. evansi, suggesting that the parasite interferes with the cholinergic neurotransmission in this experimental condition.     

Effect of diphenyl diselenide diet supplementation on oxidative stress biomarkers in two species of freshwater fish exposed to the insecticide fipronil

The ability of diphenyl diselenide [(PhSe)₂] to attenuate oxidative damage was evaluated in the liver, gills, brain, and muscle of carp (Cyprinus carpio) and silver catfish (Rhamdia quelen) experimentally exposed to fipronil (FPN). Initially, the fish were fed a diet without (PhSe)₂ or a diet containing 3.0 mg/kg of (PhSe)₂ for 60 days. After the 60-day period, the fish were exposed to 0.65 µg/L of FPN for 192 h. The results showed that carp exposed to FPN and not fed with (PhSe)₂ exhibited acetylcholinesterase (AChE) inhibition in brain and muscle, and increased thiobarbituric acid-reactive substance (TBARS) in liver, gills, and brain. Furthermore, FPN decreased nonprotein thiols (NPSH) and δ-aminolevulinate dehydratase (δ-ALA-D) in carp liver and gills, and increased plasma glucose and protein levels. In silver catfish, FPN inhibited AChE and increased TBARS levels in muscle. In addition, glutathione S-transferase (GST) decreased in liver and muscle, and plasma glucose was increased. (PhSe)₂ reversed some of these effects. It prevented the increase in TBARS levels in liver, gills, and brain in carp and in silver catfish muscle, and reversed the increase in plasma glucose levels in both species. Additionally, (PhSe)₂ increased the NPSH levels in carp and silver catfish that had decreased in response to FPN exposure. However, (PhSe)₂ was not effective in reversing the AChE inhibition in brain and muscle or the δ-ALA-D decrease in carp liver. Thus, (PhSe)₂ protects tissues of both species of fish, mainly by preventing or counteracting the effects of FPN, on TBARS levels, antioxidants, and present anti-hyperglycemic property.     

Animals selected for postweaning weight gain rate have similar maintenance energy requirements regardless of their residual feed intake classification

Data of comparative slaughter were used to determine Nellore bulls’ net energy requirements classified as efficient or inefficient according to residual feed intake (RFI) and selection lines (SL). Sixty-seven Nellore bulls from the selected (SE) and control (CO) lines of the selection program for postweaning weight gain were used. The animals underwent digestibility trials before being submitted to the finishing trial. Sixteen bulls were slaughtered at the beginning of the finishing trial, and their body composition was used as the baseline for the remaining animals. For body composition determinations, whole empty body components were weighed, ground, and subsampled for chemical analyses. Initial body composition was determined with equations developed from the baseline group using shrunk body weight, fat, and protein. The low RFI (LRFI) and CO animals had a lower dry matter (DMI) and nutrient intake (P < 0.05) than high RFI (HRFI) and SE animals, without alterations in digestibility coefficients (P > 0.05). During the finishing trial, DMI remained lower for LRFI and CO animals. Growth performance was similar between RFI classes, except for empty body weight gain that tended to be higher for LRFI than HRFI (P = 0.091). The SE animals had less fat content on the empty body (P = 0.005) than CO. Carcasses tended to be leaner for LRFI than HRFI (P = 0.080) and for SE than CO (P = 0.066) animals. LRFI animals retained more energy (P = 0.049) and had lower heat production (HP; P = 0.033) than the HRFI ones. Retained energy was not influenced by SL (P = 0.165), but HP tended to be higher for SE when compared to CO (P = 0.075) animals. Net energy requirement for maintenance (NE_m) was lower for LRFI than HRFI (P = 0.009), and higher for SE than CO (P = 0.046) animals. There was an interaction tendency between RFI and SL (P = 0.063), suggesting that NE_m was lower for LRFI+CO than HRFI+CO (P = 0.006), with no differences for SE (P = 0.527) animals. The efficiency of ME utilization for maintenance (k_m) of LRFI and HRFI animals were 62.6% and 58.4%, respectively, and for SE and CO were 59.0% and 62.1%, respectively. The breeding program for postweaning weight has not improved feed efficiency over the years, with RFI classification not being a promising selection tool for SE animals. Classification based on RFI seems to be useful in animals that have not undergone the breeding program, with LRFI animals having lower energy requirements than the HRFI ones.     

Comparison between haplotype-based and individual snp-based genomic predictions for beef fatty acid profile in Nelore cattle

The aim of this study was to evaluate the genomic predictions using the single-step genomic best linear unbiased predictor (ssGBLUP) method based on SNPs and haplotype markers associated with beef fatty acids (FAs) profile in Nelore cattle. The data set contained records from 963 Nelore bulls finished in feedlot (±90 days) and slaughtered with approximately 24 months of age. Meat samples from the Longissimus dorsi muscle were taken for FAs profile measurement. FAs were quantified by gas chromatography using a SP-2560 capillary column. Animals were genotyped with the high-density SNP panel (BovineHD BeadChip assay) containing 777,962 markers. SNPs with a minor allele frequency and a call rate lower than 0.05 and 0.90, respectively, monomorphic, located on sex chromosomes, and with unknown position were removed from the data set. After genomic quality control, a total of 469,981 SNPs and 892 samples were available for subsequent analyses. Missing genotypes were imputed and phased using the FImpute software. Haplotype blocks were defined based on linkage disequilibrium using the Haploview software. The model to estimate variance components and genetic parameters and to predict the genomic values included the random genetic additive effects, fixed effects of the contemporary group and the age at slaughter as a linear covariate. Accuracies using the haplotype-based approach ranged from 0.07 to 0.31, and those SNP-based ranged from 0.06 to 0.33. Regression coefficients ranged from 0.07 to 0.74 and from 0.08 to 1.45 using the haplotype- and SNP-based approaches, respectively. Despite the low to moderate accuracies for the genomic values, it is possible to obtain genetic progress trough selection using genomic information based either on SNPs or haplotype markers. The SNP-based approach allows less biased genomic evaluations, and it is more feasible when taking into account the computational and operational cost underlying the haplotypes inference.     

Mitoepigenetics: Methylation of mitochondrial DNA is strand-biased in bovine oocytes and embryos

Global mitochondrial DNA (mtDNA) methylation has been recently described in bovine and showed particular signatures in both gametes and embryos. Here, we investigated the distribution of mtDNA methylation through strand-specific mapping of methylation sites to gain perspective on how epigenetic mechanisms can be involved in mitochondrial function. We demonstrate that in both oocytes and embryos, the frequency of methylation is biased towards the light strand (L-strand), particularly in the gene bodies and in the region containing the L-strand promoter (LSP). Methylation is not restricted to CpG nucleotides and is not symmetrical on both strands. This configuration reinforces the hypothesis of a specific epigenetic regulation of mtDNA, which is an important observation for the understanding of how mitochondrial function is regulated.