Biofloc technology and immune response of penaeid shrimp: A meta-analysis and meta-regression.

Biological flocs develop as heterogenious communities of detritus, heterotrophic and autotrophic bacteria, algae, fungi and a variety of other things through appropriate management of the carbon-nitrogen ratio in aquaculture farming systems. These microbial communities improve the water quality, reduce the concentrations of toxic inorganic species, and act as a source of food and immunostimulants. However, the information about their role as immunostimulants is widely variable. To obtain a clearer view of the situation, it is necessary to summarize the current literature on this topic through a quantitative review analysis: a meta-analysis. The present work aims to evaluate the effect of biofloc technology (BFT) implementation on culture water quality through meta-analysis, meta-regression, and correlation analysis. Pubmed, Science Direct, Scopus, and Google Scholar were searched from 2000 to 2022 on the influence of BFT on immunological parameters. Three hundred and thirty-four of the scientific articles used BFT for penaeid shrimp, but only 25 articles met the inclusion criterion of using BFT to evaluate its effect on the shrimp's immunity. The current analysis found that the use of BFT stimulates all the evaluated immunological parameters. Also, meta-regression with fixed data of study parameters showed that slow-release carbon sources, probiotics, and an estimated carbon/nitrogen ratio of 10 potentiated a larger effect on BFT immune stimulation. The meta-analysis demonstrated that the volumes of the settleable solids (SS) and the total suspended solids (TSS) are crucial for the effect of BFT on the shrimp immune system. When TSS was higher than 294 mg/L, the respiratory metabolism of the shrimp was compromised. The analysis found evidence of publication biases, indicating that the data had to be interpreted carefully. Nonetheless, this meta-analysis shows that the data quality was sound and the approach used was appropriate; moreover, it also showed that BFT can be an environmentally friendly alternative culture method that enhances the shrimp's immune system.

Risk factors for the presence of Fasciola hepatica antibodies in bulk-milk samples and their association with milk production decreases, in Cuban dairy cattle.

BACKGROUND: Worldwide, Fasciola hepatica infection causes high production losses in the livestock industry. Recently, studies have analyzed the association between measurements of F. hepatica infection intensity and herd management practices. The aim of the present study, the first of its kind in a subtropical region, was to evaluate associations between F. hepatica bulk-tank milk ELISA results with herd management factors and milk yield in dairy herds, in Camagüey, Cuba. The SVANOVIR® F. hepatica-AB ELISA was used to measure F. hepatica antibody levels in a random sample of 516 dairy herds during the period of May-July of 2014. Farm management practice data were collected using a questionnaire. RESULTS: With 82% of the herds testing positive, the results indicate that F. hepatica is very widespread in this area. Reductions in milk production of 18 and 32% were observed in herds with Optical Density Ratios (ODR) of 0.3-0.6 and > 0.6, respectively, when compared to herds with ODR <  0.3. Overall, the longer the milking cows were put out to pasture, the higher the levels of anti-parasite antibodies. Co-grazing with sheep and goats also significantly increased the risk of high ODR. CONCLUSIONS: Our data show a widespread occurrence of the parasite as well as a major potential impact of the infection on the Cuban development goal of becoming self-sufficient in milk production. Our risk factor analysis suggests that the prevention of infection around water sources, and the separation of cattle from small ruminants could be useful control measures. This is the first epidemiological survey of F. hepatica abundance, and associated reductions in milk yield, in dairy herds in Cuba.

Use of Wing Geometric Morphometric Analysis and mtDNA to Identify Africanization of Apis mellifera in the Central Highlands of Ecuador.

Seventy-five samples were collected from 15 beehives in the central highlands of Ecuador (Tungurahua-Chimborazo) to assess Africanization in managed bee populations using wing geometric morphometric and mitochondrial DNA analyses. The results indicated that when grouping the apiaries based on altitudinal floors into 2600-2800, 2801-3000, and 3001-3274 m above sea level, differences (p < 0.001) were observed. The morphotypes were similar in the first two floors, but the third indicated that altitude plays a crucial role in the differentiation of populations. When comparing with the pure subspecies, we found differences (p < 0.001); the nearest Mahalanobis distance was for Apis mellifera scutellata (D2 = 3.51), with 95.8% Africanization via father in the area. The maternal origin of all patterns belonged to lineage A (A. m. scutellata), with seven haplotypes. The most frequent haplotypes were A26 and A1; however, the A1q haplotype was not detected at the national level or in nearby countries. The identified haplotypes do not coincide with A4, which is predominant in South Africa and Brazil. The results indicate a double origin due to their presence in North Africa and the Iberian Peninsula. The formation of specific morphological groups within ecoregions is suggested.

Geometric Morphometric Analysis of Wing Shape to Identify Populations of Apis mellifera in Camagüey, Cuba.

A total of 45 Apis mellifera colonies were sampled from nine centers for rearing queens in the Camagüey province, Cuba. Wing geometric morphometric analysis was used to determine the ancestry and identify Africanization processes at different altitudes in managed honeybee populations on the island. A total of 350 reference wings were obtained from the pure subspecies: Apis mellifera mellifera, Apis mellifera carnica, Apis mellifera ligustica, Apis mellifera caucasia, Apis mellifera iberiensis, Apis mellifera intermissa, and Apis mellifera scutellata for the study. Our results showed that altitude influences wing shape; and that 96.0% (432) of the individuals were classified as Cuban hybrids, with a tendency to the formation of a new morphotype. In addition, a great similarity was found with the subspecies Apis mellifera mellifera, and it was confirmed that there is no Africanization due to the low presence of 0.44% (2) of this morphotype in the population under study. The greatest Mahalanobis distances were obtained for the comparisons between the center rearing of queens in the Camagüey province with the subspecies A. m. scutellata (D2 = 5.18); A. m. caucasia (D2 = 6.08); A. m. ligustica (D2 = 6.27); and A. m. carnica (D2 = 6.62). The well-defined pattern of wing shape produced by honeybee populations in Camagüey's centers for queen rearing suggests a Cuban hybrid. Moreover, it is essential to note that the populations of bees under investigation lack Africanized morphotypes, indicating that Camagüey bees have not interacted with the African lineage.

Hygienic Behavior of Apis mellifera and Its Relationship with Varroa destructor Infestation and Honey Production in the Central Highlands of Ecuador.

The aim of this research was to analyze the relationship among hygienic behavior (HB), Varroa destructor infestation, and honey production in the central highlands of Ecuador. Overall, 75 honey bee colonies were evaluated before, during, and after production at three altitude levels (2600-2800, 2801-3000, and >3000 m.a.s.l.). The hygienic behavior percentage of the colonies was determined by the pin-killing method, and the colonies were classified into three groups: high HB (>85%), mid HB (60.1-85%), and low HB (≤60%). Varroa infestation was diagnosed as well, and honey production was evaluated only during production. HB was high and heterogeneous, averaging 80% ± 9.7%. Its highest expression was observed at lower altitudes. The infestation degree was low (3.47% ± 1.56%), although the mite was detected in all colonies upon sampling. A negative correlation was observed between HB and Varroa infestation in the first sampling (-0.49 **), suggesting that the high- and mid-altitude HB colonies underwent the lowest infestation rates, regardless of sampling. The correlations between HB and production were significant (0.26 *), indicating a positive effect of HB on production, meaning that colonies with high HB obtained the highest honey production (25.08 ± 4.82 kg/hive). The HB of bees showed an inverse relationship with altitude and it tended to reduce the effect of Varroa infestation, favoring honey production and, thus, suggesting the feasibility of selecting colonies with high HB.

Altered non-coding RNA expression profile in F1 progeny 1 year after parental irradiation is linked to adverse effects in zebrafish.

Gamma radiation produces DNA instability and impaired phenotype. Previously, we observed negative effects on phenotype, DNA methylation, and gene expression profiles, in offspring of zebrafish exposed to gamma radiation during gametogenesis. We hypothesize that previously observed effects are accompanied with changes in the expression profile of non-coding RNAs, inherited by next generations. Non-coding RNA expression profile was analysed in F1 offspring (5.5 h post-fertilization) by high-throughput sequencing 1 year after parental irradiation (8.7 mGy/h, 5.2 Gy total dose). Using our previous F1-γ genome-wide gene expression data (GSE98539), hundreds of mRNAs were predicted as targets of differentially expressed (DE) miRNAs, involved in pathways such as insulin receptor, NFkB and PTEN signalling, linking to apoptosis and cancer. snRNAs belonging to the five major spliceosomal snRNAs were down-regulated in the F1-γ group, Indicating transcriptional and post-transcriptional alterations. In addition, DEpiRNA clusters were associated to 9 transposable elements (TEs) (LTR, LINE, and TIR) (p = 0.0024), probable as a response to the activation of these TEs. Moreover, the expression of the lincRNAs malat-1, and several others was altered in the offspring F1, in concordance with previously observed phenotypical alterations. In conclusion, our results demonstrate diverse gamma radiation-induced alterations in the ncRNA profiles of F1 offspring observable 1 year after parental irradiation.

Growth enhancement of shrimp (Litopenaeus schmitti) after transfer of tilapia growth hormone gene.

Electroporation of Litopenaeus schmitti embryos was used to transfer the pE300tiGH15 plasmid that contains the tilapia growth hormone gene (tiGH) complexed with a nuclear localization signal peptide into the zygotes. The gene construct was detected in 35 (36%) of the 98 larvae screened by PCR and Southern blot analyses. Western blot analyses revealed that 34% of the screened larvae expressed a single tiGH-specific band with the expected molecular mass (23.1 kDa). The development index and larval length indicated a significant growth enhancement from day 3 on after electroporation, with an average of 32% of the growth enhancement. To our knowledge, this is the first report on gene transfer enhanced growth in crustaceans.

Bovine fascioliasis in Brazil: Economic impact and forecasting.

Fasciola hepatica causes liver damage and poor growth in cattle and other animals, including humans. Although the disease occurs throughout the country, it is hyperendemic in cattle in the South of Brazil. This work aimed to determine the economical loss by carcass weight variance using data from all the states where F. hepatica is found, as well as to run a disease forecast analysis for the Rio Grande do Sul State. We found a direct loss of approximately US$ 210 million/year from infected cattle in Brazil and the ARIMA model analysis revealed that an increase of fascioliasis is most probable if no parasite control program is adopted.

Growth Hormone Secretagogue (A233) Improves Growth and Changes the Tissue Fatty Acid Profile in Juvenile Tilapia (Oreochromis niloticus).

Growth hormone (GH) release is a process that is well regulated by several factors, including GH secretagogues. GH can mediate the regulation of the fatty acid level and composition. The aim of this study was to determine the effect of a synthetic GH secretagogue peptide (A233) on the growth and fatty acid composition in tilapia (Oreochromis niloticus). To address this objective, we administrated a diet supplemented with A233 to juvenile tilapia for 60 days. The group fed with a diet supplemented with 600 µg of A233 per kg of feed increased in weight (4.81 ± 0.09 g) and specific growth rate (2.49 ± 0.03%/day) compared to the control diet group (3.63 ± 0.08 g, 2.07 ± 0.04%/day; respectively) (p < 0.001). In the muscle, the total lipids for the control diet group were higher than that in the group fed with 600 µg of A233 per kg feed; however, no differences were detected in the liver. In both tissues, the patterns of fatty acid composition and content were generally similar, with some exceptions. Tilapia fed with 600 µg of A233 per kg of feed showed, in liver and muscle, a significantly higher composition and content of n-3 polyunsaturated fatty acids (such as 20:5n-3, 22:5n-3, 22:6n-3) and n-3/n-6 PUFA than animals fed with the control diet. To our knowledge, this is the first report on the the effects of natural or synthetic GH secretagogues (GHS) on fatty acid composition, implying an increase in the nutritional quality of the tilapia.

Sheep polyclonal antibody to map Haemonchus contortus mimotopes using phage display library.

The aim of this study was to evaluate phage display technology for mapping Haemonchus contortus mimotopes. We screened the PhD-7 Phage Display Peptide Library Kit with a sheep polyclonal antibody against H. contortus. After four rounds of selection, 50 phage peptide clones were selected by biopanning and sequenced. Two clones displaying peptide mimotopes of H. contortus proteins were chosen for sheep immunization: clone 6 - mimotope of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and clone 17 - mimotope of a disorganized muscle family member (Dim 1). Twelve sheep were allocated into 3 groups of 4 animals as follow: G1: control group; G2/GAPDH: immunized with clone 6; and G3/Dim1: immunized with clone 17. Four immunizations were performed at intervals of seven days (0, 7, 14, and 21 days). On day 28 post initial vaccination, all groups were orally challenged with 2500 H. contortus infective larvae. The mimotope peptides selected by phage display were recognized by IgG from sheep naturaly infected with H. contortus. The immunization protocol showed an increasein IgG anti-M13 phage titers, but no effect was observed in IgG-specific for the anti-mimotope peptides. This is the first report of successful use of a phage display library for the identification of mimotopes of H. contortus proteins.

Diagnosis of resistance alleles in codon 167 of the beta-tubulin (Cya-tbb-1) gene from third-stage larvae of horse cyathostomins.

Anthelmintic resistance is a serious problem for the control of equine gastrointestinal nematodes. In the present survey, 173 third stage larvae of cyathostomins were investigated from three different locations for the presence of the resistant genotype at codon 167 of the beta-tubulin gene, as this is the most prevalent mutation. The larvae from the state of Parana (n=67), Sao Paulo (n=54) and Santa Catarina (n=52), showed 61.2; 31.5 and 38.5% of the heterozygous resistant genotype - TTC/TAC, respectively. An unpublished mutation at codon 172 that results in a serine (S) to threonine (T) substitution was found in 17.9% (12/67) of samples from Parana; and 13.0% (7/54) of samples from Sao Paulo. We have compared the molecular diagnostic with the fecal egg count data (R2=-0.79) from the same farms, and consider that the use of routine molecular diagnostic in individual larva may help to determine the population genetic distribution that is associated with drug failure.

Efficacy of two extra-label anthelmintic formulations against equine strongyles in Cuba.

Equine cyathostomin parasites are ubiquitous in grazing horses and have been shown to cause severe inflammatory disease in the large intestine of horses. Decades of intensive anthelmintic therapy have led to widespread anthelmintic resistance in cyathostomins across the world. In Cuba, no anthelmintic products are formulated and sold for equine usage and little is known about anthelmintic efficacy of ruminant and swine formulations used. A strongyle fecal egg count reduction test was used to assess the efficacy of a liquid formulation of ivermectin labelled for use in swine, ruminants and carnivores and a pelleted formulation of albendazole labelled for usage in ruminants. Nine farms in the province Camagüey were enrolled in the study comprising 149 horses in total. Albendazole efficacy was reduced on five farms and with the other four farms having no signs of reduced efficacy. Mean farm efficacies were ranging from 41.7% to 100% on the tested farms. Coprocultures found large strongyle larvae present on all farms, but all larvae identified post-treatment were cyathostomins. Ivermectin was found 100% efficacious on all studied farms. This study provided evidence of reduced albendazole efficacy in the study population. Further work is needed to evaluate whether these findings reflect true resistance or if they are due to pharmacokinetic or pharmacodynamic characteristics of the pelleted formulation tested here.

Gene silencing of Dim-1, a member of the disorganized muscle family, in Haemonchus contortus.

RNA interference has been widely used in parasites. In Haemonchus contortus, reproducible silencing has been reported; however, in this species an altered phenotype has been observed for only a few genes silenced by RNA interference. The aim of this work was to evaluate the effect of silencing Dim-1, a member of the disorganized muscle family on third stage larvae (L3) of H. contortus. Effective silencing of Dim-1 in L3 led to reduced L3 migration and slowed larval development from L3 to early L4. To our knowledge, this is the first report to demonstrate Dim-1 silencing affecting phenotypic characteristics of this parasite.

The SV40 T antigen nuclear localization sequence enhances nuclear import of vector DNA in embryos of a crustacean (Litopenaeus schmitti).

A genetic transformation system for penaeid shrimp could provide a powerful technique for the improvement of different production traits of importance for a sustainable aquaculture. The development of a successful transformation system depends on the ability to efficiently introduce exogenous DNA into the target species. The ability of the nuclear localization signal (NLS) peptide of the SV40 T antigen to facilitate nuclear import and transient gene expression is known from vertebrate systems and for the first time, is shown here to be efficient in a crustacean species, i.e. the shrimp Litopenaeus schmitti. Electroporation was used to introduce the pCMV-lacZ plasmid that contains the human cytomegalovirus promoter/enhancer (CMV) fused to the beta-galactosidase (lacZ) coding region, into L. schmitti zygotes. Supercoiled DNA was used at 50 or 500 ng/microl naked or bound to NLS peptide. The hatching rate of electroporated zygotes was around 60% for all groups, except from the pCMV-lacZ:NLS group at 500 ng/microl (43%). Based on Southern blot analyses of polymerase chain reaction (PCR) products the gene transfer frequency was 2-fold higher using DNA:NLS complexes than with naked DNA (23.8% vs. 11.5%, with 50 ng/microl of plasmid DNA, 44.3% vs. 28.8% with 500 ng/microl). The beta-galactosidase activity assay indicated that nuclear uptake is faster for the DNA:NLS complexes than for naked DNA. The beta-galactosidase activity was always higher in the DNA:NLS groups than in the naked DNA groups. To our knowledge, this is the first report on the use of an NLS peptide to improve gene transfer and nuclear uptake in crustaceans.

Sheep polyclonal antibody to map Haemonchus contortus mimotopes using phage display library / Anticorpo policlonal de ovinos para mapear mimetopos de Haemonchus contortus usando a biblioteca de Phage display

Abstract The aim of this study was to evaluate phage display technology for mapping Haemonchus contortus mimotopes. We screened the PhD-7 Phage Display Peptide Library Kit with a sheep polyclonal antibody against H. contortus. After four rounds of selection, 50 phage peptide clones were selected by biopanning and sequenced. Two clones displaying peptide mimotopes of H. contortus proteins were chosen for sheep immunization: clone 6 - mimotope of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and clone 17 - mimotope of a disorganized muscle family member (Dim 1). Twelve sheep were allocated into 3 groups of 4 animals as follow: G1: control group; G2/GAPDH: immunized with clone 6; and G3/Dim1: immunized with clone 17. Four immunizations were performed at intervals of seven days (0, 7, 14, and 21 days). On day 28 post initial vaccination, all groups were orally challenged with 2500 H. contortus infective larvae. The mimotope peptides selected by phage display were recognized by IgG from sheep naturaly infected with H. contortus. The immunization protocol showed an increasein IgG anti-M13 phage titers, but no effect was observed in IgG-specific for the anti-mimotope peptides. This is the first report of successful use of a phage display library for the identification of mimotopes of H. contortus proteins.

Resumo O objetivo deste estudo foi avaliar a tecnologia de phage display no mapeamento de mimetopos de Haemonchus contortus. Anticorpo policlonal de ovinos anti-H. contortus foi usado para seleção a partir da biblioteca PhD-7 Phage Display Peptide Library Kit (New England BioLabs). Após quatro rodadas, 50 clones de fagos expressando peptídeos foram selecionados e sequenciados. Dois clones que exibiram mimetopos de H. contortus foram escolhidos para imunização de ovinos: clone 6 - mimetopo de gliceraldeído-3-fosfato desidrogenase (GAPDH) e clone 17 - mimetopo da família do músculo desorganizado (Dim 1). Doze ovinos foram alocados em 3 grupos de 4 animais, da seguinte forma: G1: grupo controle, G2/GAPDH: imunizado com o clone 6 e G3/Dim1: imunizado com o clone 17. Quatro imunizações foram realizadas (0, 7, 14 e 21 dias). No dia 28 após a primeira imunização, todos os grupos foram desafiados oralmente com 2500 larvas infectantes de H. contortus . Os peptídeos mimetopos selecionados foram reconhecidos por IgG de ovinos naturalmente infectados por H. contortus. O ensaio de imunização revelou um aument dos títulos de IgG anti-fago M13, mas não ocorreu aumento de IgG anti-peptídeos mimetopos. Este é o primeiro relato de uso bem sucedido da biblioteca de Phage display para a identificação de mimetopos de H. contortus.

Aqueous extract of Ocimum tenuiflorum decreases levels of blood glucose in induced hyperglycemic tilapia (Oreochromis niloticus).

OBJECTIVE: To evaluate, in hyperglycemic tilapia [Oreochromis niloticus (O. niloticus)], the effect of this aqueous extract on blood glucose levels. METHODS: The hyperglycemia in O. niloticus was induced by adding glucose to fish pond water. An aqueous extract of Ocimum tenuiflorum (O. tenuiflorum) was prepared by boiling fresh leaves and the doses of 0, 40, 80, 200 and 400 mg per liter of pond water were tested. RESULTS: The blood sugar concentration for tilapia with hyperglycemic induced was an average of 50% higher than the control group. The blood glucose levels in tilapia after the induction of hyperglycemia were higher than the control group for 90 min after the treatment. The treatment with the aqueous extract of O. tenuiflorum dropped the serum glucose level of hyperglycemic tilapia until it was similar to that of the control group and was dose dependent. CONCLUSIONS: The results indicated that O. tenuiflorum was endowed with anti-hyperglycemic activity. To our knowledge, this is the first report on the use of fish as a diabetes model to test natural extracts from plants.