Applications of livestock monitoring devices and machine learning algorithms in animal production and reproduction: an overview

Some sectors of animal production and reproduction have shown great technological advances due to the development of research areas such as Precision Livestock Farming (PLF). PLF is an innovative approach that allows animals to be monitored, through the adoption of cutting-edge technologies that continuously collect real-time data by combining the use of sensors with advanced algorithms to provide decision tools for farmers. Artificial Intelligence (AI) is a field that merges computer science and large datasets to create expert systems that are able to generate predictions and classifications similarly to human intelligence. In a simplified manner, Machine Learning (ML) is a branch of AI, and can be considered as a broader field that encompasses Deep Learning (DL, a Neural Network formed by at least three layers), generating a hierarchy of subsets formed by AI, ML and DL, respectively. Both ML and DL provide innovative methods for analyzing data, especially beneficial for large datasets commonly found in livestock-related activities. These approaches enable the extraction of valuable insights to address issues related to behavior, health, reproduction, production, and the environment, facilitating informed decision-making. In order to create the referred technologies, studies generally go through five steps involving data processing: acquisition, transferring, storage, analysis and delivery of results. Although the data collection and analysis steps are usually thoroughly reported by the scientific community, a good execution of each step is essential to achieve good and credible results, which impacts the degree of acceptance of the proposed technologies in real life practical circumstances. In this context, the present work aims to describe an overview of the current implementations of ML/DL in livestock reproduction and production, as well to identify potential challenges and critical points in each of the five steps mentioned, which can affect results and application of AI techniques by farmers in practical situations.(AU)

Use of new recombinant proteins for ovarian stimulation in ruminants

Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.(AU)

Gestational follow-up in ewes - placentome size and fecal progesterone metabolite levels

Background: Transabdominal ultrasonography in sheep allows real-time diagnosis and monitoring of pregnancy as well as measurement of the size of placentomes, which are structures that connect the fetal and maternal portions, that increase as pregnancy progresses. Progesterone is involved in the maintenance of gestation, with high levels noted during pregnancy. In this context, it is hypothesized that measurements of placentome and progesterone could help in monitoring pregnancy in sheep. Therefore, the main objectives of this study included evaluating whether placentome size and fecal progesterone levels are associated with gestational age in ewes and whether measurement of the placentome and progesterone could be used as an estimate of gestational age. Materials, Methods & Results: A total of 63 pregnant adult (between 2 and 5 years of age) Santa Inês ewes were monitored in the city of Boa Vista, Roraima. All ewes were kept in the paddock with Panicum maximun cv. tanzania, mineral salt, and water ad libitum, supplemented with 200 g/animal/day of soybean residue. Ewes were subjected to ultrasonography every 21 days from day 42 of pregancy until term to monitor pregnancy. Ultrasonography was performed using a linear probe to measure the placentome, including length × width in centimeters. Also every 21 days, fecal samples were collected for measurement of progesterone level. Feces were collected directly from the rectal ampulla, stored individually in plastic bags, and frozen at -20ºC until processing. In the laboratory, these feces samples were freeze-dried and subjected to hormonal extraction with 80% methanol. Then, the levels of fecal progesterone metabolites were measured by enzyme immunoassay. Data was tabulated and submitted to statistical analyses, including descriptive stastistic, correlation, KruskalWallis test and Tukey's test. The levels of fecal progesterone metabolites increased significantly between the 3rd and 4th months of pregnancy, with a decrease close to delivery. Placentome size increased significantly between the 2nd and 3rd months of pregnancy. Placentome measurements did not correlate with progesterone levels and showed a weak correlation with gestational age. Therefore, we concluded that the measurements of placentome size and a single evaluation of fecal progesterone metabolites were not considered efficient methods for estimating gestational age. Discussion: Measurement of progesterone level in fecal samples is a non-invasive hormone monitoring method that provides values like that of blood levels and is less stressful for the animals at the time of sample collection. The wide physiological variation makes it difficult or even useless to use fecal progesterone, when is measured only once per animal, as an indicator of the presence of pregnancy. Despite this limitation, serial sampling is useful for monitoring of hormones during the pregnancy period and is an important tool for research purposes. Placentome size measurements correlated weakly with gestational age and did not correlate with progesterone level. Thus, the presence of placentomes is indicative of the presence of pregnancy but was considered inefficient for determining gestational age in sheep.

Essential fatty acids in farmed tambaqui (Colossoma macropomum) from the Brazilian Amazon Area

The goal of this study was to determine the essential fatty acids of the total lipids of the fillet, head and orbital cavity tissue from farmed tambaqui (Colossoma macropomum) fish from a Brazilian Amazon area. The tambaqui were acquired from different fish farms in the Roraima state, located at Western Brazilian Amazon. The meat, the head and the fatty tissue from orbital cavity were dissected for lipid extraction and analysis of fatty acids by gas chromatography. The fatty acids were quantified in mg g-1of total lipids using C23:0 as an internal standard. The nutritional quality of the lipids was determined by using the atherogenicity and thrombogenicity indices, and also by the ratio between hypocholesterolemic / hypercholesterolemic fatty acids. The orbital cavity tissue had the higher concentration amount of linoleic and α-linolenic acid, whereas the fillet had higher docosahexaenoic acid (DHA). The eicosapentaenoic acid (EPA) concentration was: 1.28, 0.97, 1.71 mg g-1of total lipids, in the filet, in head, and in orbital cavity tissue,respectively. All essential fatty acids were detected in the three parts analyzed. The nutritional quality of the total lipids from the head and from the orbital cavity tissue was similar to the fillet.(AU)

Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria / Eugenia uniflora: uma alternativa natural promissora contra bactérias multirresistentes

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Este trabalho teve como objetivo avaliar a composição química, as atividades antioxidantes e antimicrobianas do extrato bruto e frações de folhas de Eugenia uniflora Linn. O extrato bruto foi obtido por turbólise e suas frações por partição. Foram realizadas análises cromatográficas e a capacidade antioxidante foi verificada por dois métodos (DPPH• e ABTS•+). A Concentração Inibitória/Bactericida Mínima foi realizada contra vinte e duas bactérias, selecionando cinco cepas suscetíveis a extração/frações e resistentes aos antibióticos testados. Ampicilina, azitromicina, ciprofloxacina e gentamicina foram associados à Fração Acetato de Etila (FAE) contra cepas multirresistentes em testes modulatórios e de checkboard. Os dados cromatográficos mostraram ácido gálico, ácido elágico e miricitrina em extrato bruto, com enriquecimento na FAE. A atividade de transferência de elétrons demonstrada nos testes antioxidantes está relacionada com a presença de flavonoides. As cepas de Gram-positivas foram mais suscetíveis à FAE, e seus espectros de ação foram melhorados por associação, compreendendo bacilos Gram-negativos. Foram observados sinergismos de ciprofloxacina e gentamicina contra Pseudomonas aeruginosa resistente à colistina. Os resultados demonstram que o extrato e a fração enriquecida obtida das folhas de E. uniflora atuam como uma alternativa natural promissora contra bactérias multirresistentes.

Cryptosporidium spp. in dogs - prevalence and genotype distribution

Background: Cryptosporidium spp. is a zoonotic protozoan parasite that affects the gastrointestinal tract of humans and animals. The disease can cause acute and chronic diarrhoea and even death in both humans and animals. In this study, it was aimed to determine the prevalence and genotype distribution of Cryptosporidiosis in shelter dogs in Diyarbakir province located in the Southeastern Anatolia Region of Turkey. Materials, Methods & Results: The animal material of the study consisted of 100 dogs of different breeds and sexes. Faecal samples were collected from the rectum with disposable latex gloves and placed in individual sample containers. All of the samples were examined for Cryptosporidium spp. by Kinyoun Acid Fast and Nested PCR methods. In the Kinyoun Acid Fast staining method, firstly, smear preparations were prepared from fresh faecal samples, fixed in pure methanol for 1 min and allowed to dry. The slides were kept in Kinyoun Carbol-Fuxin for 5 min, dipped in 50% ethyl alcohol, shaken, washed in tap water, kept in 1% sulphuric acid for 2 min and washed in tap water. The slides were kept in methylene blue for 1 min, washed in tap water and allowed to dry. After drying, immersion oil was dripped and examined under a microscope at 100 magnification. DNA extraction was performed from all samples using GeneMATRIX Stool DNA Purification Kit according to the manufacturer's protocol. After Nested PCR analysis was performed. In the PCR step, primers 5'-TTCTAGAGCTAATACATGCG-3' and 5'- CCCATTTCCTTCCTTCGAAACAGGA-3' were used to amplify the 1325 bp gene region. In the nested PCR step, primers 5'- GGAAGGGTTGTATTTATTTATTAGATAAAG-3' and 5'-AAGGAGTAAGGAACAACCTCCA-3' were used to amplify the 826-864 bp gene region. As a result of both methods, a prevalence of 3% was determined. The infection rate was higher in males (3.57%) than females (2.27%) and in younger than 1 year (5.56%) than in older than 1 year (1.56%). The DNA sequences obtained from the sequence analysis of 3 positive PCR samples were analysed in BioEdit software. A phylogenetic tree was constructed with the data set created by using the 18s rRNA gene sequences obtained from the NCBI genbank database and the DNA sequences obtained as a result of the study, and it was shown which Cryptosporidium species the study samples were related to. Today, many Cryptosporidium species have been identified and most of these species have host adaptation. Although C. canis is the most common species in dogs, C. muris, C. meleagridis, and C. parvum have also been detected. Among these species, C. parvum is recognized as a zoonotic species infecting a wide range of mammals. In this study, DNA sequencing of nested PCR positive samples revealed that 3 samples were zoonotic C. parvum. Discussion: This suggests that dogs may be a reservoir for zoonotic transmission of Cryptosporidium. Consequently, it is recommended that people should be informed about the potential for transmission of this protozoan to humans and animals and that control programmes should be implemented, including the prevention of free entry of stray dogs into public places and homes.

Potassium extraction from the silicate rock Verdete using organic acids

Although Brazil is one of the world’s leading exporter of agricultural products, the country is highly dependent on the importation of potassic fertilizers. K-bearing silicate rocks are reported as potential solutions to reduce external K dependency. This work evaluated K extraction from silicate Verdete rock, a glauconite-bearing rock containing 10 % of K2O, by solubilization with organic acids. Firstly, Verdete rock was reacted during 3-120 h with solutions of citric or oxalic acid at 2 % (m/v) in Erlenmeyer flasks by shaking. Oxalic acid extracted 6.5 % of K in Verdete, while citric acid extracted 2.3 %. Another experiment was performed to evaluate the effect of various oxalic acid concentrations (2, 4, 6, 8, and 10 %) and differing reaction times (12, 24, 48, and 72 h) on K extraction from Verdete rock. Soluble K concentration nearly doubled with the increase of reaction time from 12 to 72 h, rising from 20 to 37 mg L–¹. Increments in K extraction were obtained by increasing oxalic acid concentrations up to 6 % and above this concentration, no significant gain was observed. The X-ray diffraction data showed that K extraction resulted from the formation of oxalate-metal complexes with metals in Verdete rock.

Potassium extraction from the silicate rock Verdete using organic acids

Although Brazil is one of the world's leading exporter of agricultural products, the country is highly dependent on the importation of potassic fertilizers. K-bearing silicate rocks are reported as potential solutions to reduce external K dependency. This work evaluated K extraction from silicate Verdete rock, a glauconite-bearing rock containing 10 % of K2O, by solubilization with organic acids. Firstly, Verdete rock was reacted during 3-120 h with solutions of citric or oxalic acid at 2 % (m/v) in Erlenmeyer flasks by shaking. Oxalic acid extracted 6.5 % of K in Verdete, while citric acid extracted 2.3 %. Another experiment was performed to evaluate the effect of various oxalic acid concentrations (2, 4, 6, 8, and 10 %) and differing reaction times (12, 24, 48, and 72 h) on K extraction from Verdete rock. Soluble K concentration nearly doubled with the increase of reaction time from 12 to 72 h, rising from 20 to 37 mg L-1. Increments in K extraction were obtained by increasing oxalic acid concentrations up to 6 % and above this concentration, no significant gain was observed. The X-ray diffraction data showed that K extraction resulted from the formation of oxalate-metal complexes with metals in Verdete rock.

Determination of lipids and fatty acids in infant formula: comparison of the conventional x direct method / Lipídios e ácidos graxos em fórmula infantil: comparação de metodologias de extração convencional x direta

Lipids in food are conventionally analyzed in two stages: extraction with organic solvent and fat esterification reaction, in this case, the type of fat of each food influences the choice of extraction and esterification reagents. In the direct method, such procedures are performed in one step. This work compared the conventional extraction method and quantification of lipids and fatty acids, with a direct method in infant formula. A reference sample of infant formula conteining certified lipids and fatty acids values from the National Institute of Standards and Technology was used. The conventional method for lipid analysis used the acid hydrolysis method; for the determination of fatty acids, the fats were extracted with a mixture of ethyl ether and petroleum ether. The direct method consisted of direct trans esterification with sodium methoxide. In the analysis of fatty acids, the majority of the results showed statistically equal values (α < 0.05) for both methods. The direct method proved suitable, mainly because of reduction in analytical time and quantity of solvents (AU).

Os lipídios em alimentos são analisados, de forma convencional, em duas etapas: extração com solvente orgânico e reação de esterificação da gordura, neste caso o tipo de gordura de cada alimento influencia na escolha dos reagentes da extração e esterificação. No método direto, estes procedimentos são realizados em uma etapa única. Este trabalho comparou a metodologia convencional de extração e quantificação de lipídios e ácidos graxos, com um método direto em fórmula infantil. Foi utilizada uma amostra de referência de fórmula infantil com valores certificados para lipídios e ácidos graxos da Nacional Institute of Standards and Technology. A metodologia convencional para a análise de lipídios empregou método com hidrólise ácida; para a determinação dos ácidos graxos, a gordura foi extraída com uma mistura de éter etílico e éter de petróleo. O método direto fundamentou-se na transesterificação direta com metóxido de sódio. Na análise dos ácidos graxos, a maioria dos resultados demonstrou valores estatisticamente iguais (α < 0,05) para os dois métodos. O método direto demonstrou ser apropriado, principalmente pela diminuição do tempo de análise e quantidade de solventes (AU).

Standardization of molecular techniques for the detection and characterization of intestinal protozoa and other pathogens in humans

Background: The intrinsic sensitivity limitations of basic parasitological methods, along with the particular biological characteristics of parasites, make these methods ineffective to differentiate morphologically indistinguishable species. Molecular detection and characterization techniques could be used to overcome these problems. The purpose of this work was to standardize molecular polymerase chain reaction (PCR) techniques, described in the literature, for the detection and molecular characterization of intestinal protozoa and other pathogens in humans. Methods: DNA was extracted from human or animal feces, previously washed or cultured in Boeck Drbohlav's Modified Medium. DNA extraction was performed with Machery-Nagel extraction kits. The standardization of the PCR, nested-PCR or RFLP techniques was carried out according to the literature. For each molecular technique performed, the sensitivity of the test was determined based on the minimun quantity required of DNA (sensitivity A) and the minimum quantity of life forms that the test detected (sensitivity B). Results: Sensitivity A was 10 fg for G. duodenalis, 12.5 pg for Entamoeba histolytica or Entamoeba dispar, 50 fg for Cryptosporidium spp., 225 pg for Cyclospora spp. and 800 fg or 8 fg for Blastocystis spp. after performing a 1780 bp PCR or 310 bp nested PCR, respectively. The sensitivity B was 100 cysts for G. duodenalis, 500 cysts for E. histolytica or E. dispar, 1000 oocysts for Cyclospora spp. and 3600 or four vegetatives forms for PCR or nested PCR of Blastocystis spp., respectively. Conclusions: The molecular detection of protozoa and chromist was achieved and the molecular characterization allowed the genotyping of some of the parasites such as Giardia duodenalis, Cryptosporidium spp., and Blastocystis spp. This study summarizes the molecular techniques for epidemiological studies in humans and animals, and helps in the investigation of their transmission sources in countries where intestinal parasites are a public health problem.(AU)

Sampling techniques and environmental variables influence the distribution of pseudoscorpions in urban forest fragments in the central Amazon / Métodos de amostragem e variáveis ambientais influenciam a distribuição de pseudoescorpiões em fragmentos florestais urbanos na Amazônia central

The growth of urban landscapes has genarally reduced biodiversity worldwide. Invertebrates have explored different environments, and it usually takes different sampling techniques to get a representative sample of the species assemblage in a given location. Some studies have sought to determine the minimum necessary number of sampling techniques, including ecological relationships or costs to guide the sampling protocol. In the Amazon, the effect of soil characteristics on invertebrate distribution is well known. We evaluated if sampling techniques have a complementary effect on the detection of pseudoscorpion assemblages and tested whether environmental variables affect the distribution of pseudoscorpion species. The study sites were two urban forest fragments in the city of Manaus, in the central Amazon. In each fragment, we sampled 20 palm trees using the beating technique, and installed transects with 12 sampling points for collection of soil and litter samples for extraction of arthropods in a Berlese funnel and a Winkler extractor, respectively. We collected 267 individuals of 11 species of pseudoscorpions. Most records were obtained through the Winkler extraction in both fragments. The assemblage from the palm trees was different from that in the edaphic samples. Pseudoscorpion species composition also differed significantly between soil and litter, and was influenced by potassium concentration. The number of species in the fragments and the environmental effect on the distribution of pseudoscorpions was similar to that recorded in environmental protection areas, evidencing that urban forest fragments can serve as an efficient repository of Amazonian pseudoscorpion biodiversity.(AU)

O crescimento das paisagens urbanas geralmente reduziu a biodiversidade em todo o mundo. Os invertebrados exploram diferentes ambientes, e geralmente são necessárias diferentes técnicas de amostragem para obter uma amostra representativa da assembleia de espécies em um determinado local. Alguns estudos têm buscado determinar o número mínimo necessário de técnicas de amostragem, incluindo relações ecológicas ou custos para orientar o protocolo de amostragem. Na Amazônia, o efeito das características do solo na distribuição dos invertebrados é bem conhecido. Nós avaliamos se as técnicas de amostragem têm um efeito complementar na detecção de assembleias de pseudoescorpiões e testamos se as variáveis ambientais afetam a distribuição das espécies de pseudoescorpiões. Os locais de estudo foram dois fragmentos florestais urbanos na cidade de Manaus, Amazônia central. Em cada fragmento, amostramos 20 palmeiras com guarda-chuva entomológico e instalamos transectos com 12 pontos amostrais para coleta de solo e serrapilheira para extração de artrópodes em funil de Berlese e extrator de Winkler, respectivamente. Coletamos 267 indivíduos de 11 espécies de pseudoescorpiões. A maioria dos registros foi obtida com Winkler em ambos fragmentos. A assembleia em palmeiras foi diferente das amostras edáficas. A composição de espécies de pseudoescorpiões também diferiu significativamente entre solo e serapilheira, e foi influenciada pela concentração de potássio. O número de espécies nos fragmentos e o efeito ambiental na distribuição de pseudoescorpiões foram semelhantes aos registrados em áreas de proteção ambiental, evidenciando que fragmentos florestais urbanos podem servir como um eficiente repositório da biodiversidade de pseudoescorpiões amazônicos.(AU)

Chemical composition and antioxidant activity of leaf essential oil from Calyptranthes concinna DC. (Myrtaceae)

The biodiversity found in Brazilian's ecosystems brings the possibility of discovering new natural products with wide application potentials. However, knowing their availability and chemical composition is crucial. Thus, the aim of this study was to evaluate the extraction yield, chemical composition and antioxidant activity of essential oil from fresh leaves of Calyptranthes concinnaDC., a native species of Myrtaceae occurring in Brazilian Atlantic Rain Forest. Plant samples were collected in SoutheasternBrazil and the essential oil was extracted by hydrodistillation. The chemical composition was evaluated by Gas Chromatography associated with Mass Spectrometry and antioxidant activity was measured using ABTS, DPPH and FRAP methods. The extraction yield obtained was 0.015% (v), and the chemical composition revealed elemicin, a phenylpropanoid as the major component (36.46%). Still, ß-caryophyllene (16.94%), germacrene B (8.28%) and spathulenol (7.33%) proved to be relevant for the same essential oil. Antioxidant activity was obtained for ABTS and DPPH radical scavenge (134.82 ± 2,9 and 93.70 ± 1.7 µM TE mL-1, respectively) and FRAP (11.31 ± 0.2 µM FeSO4mL-1OE), revealing hydrogen-donation as the main antioxidant mechanism. To our knowledge, this is the first report of antioxidant activity of C. concinna essential oil. The product presented compounds of great relevance, with possibilities of application in different areas including food, agriculture and pharmaceutical segments(AU)

Utilização de resíduos de frutos na produção e composição nutricional da larva de Tenebrio molitor / Use of fruit residues in the production and nutritiona l composition of the larva of Tenebrio Molito larva

Este estudo teve como objetivo avaliar a utilização de resíduos de frutos típicos brasileiros sobre o perfil nutricional da larva de Tenebrio molitor. O delineamento experimental foi o bloco casualizado, onde foram testadas quatro dietas diferentes para o desenvolvimento de larvas do T. molitor. O tratamento controle foi composto de trigo, soja e milho, enquanto os outros três tratamentos consistiram em dietas suplementadas com 50% de subprodutos da extração de macaúba, buriti e cacau. Foram realizados quatro tratamentos com quatro repetições entre maio e junho de 2019. Os dados foram analisados através dos testes ANOVA e Holm-Sidak, onde valores de p<0,05 foram considerados estatisticamente significativos. As larvas de T. molitor completaram seu ciclo de vida em todas as dietas testadas, entretanto, na dieta suplementada com cacau, o desenvolvimento em número de larvas foi reduzido em relação aos outros grupos. A farinha dos animais alimentados com a dieta à base de cacau apresentou um teor de proteínas inferior quando comparada à do ao grupo controle (p<0,001). Dietas à base de buriti, cacau e macaúba apresentaram quantidades maiores de extrato etéreo (p<0,018, <0,012 e <0,003, respectivamente). Concluiu-se que a suplementação de Tenébrios com dietas à base de buriti e macaúba pode ser uma boa estratégia para a melhoria do perfil nutricional da farinha de T. molitor, fornecendo quantidades favoráveis de proteínas e minerais. No tocante ao uso do cacau, estudos futuros para avaliar a presença de componentes alcaloides e seus efeitos diretos na suplementação animal devem ser conduzidos, o que pode representar uma limitação deste estudo.

This study aimed to evaluate the use of typical Brazilian fruit residues on the nutritional profile of the Tenebrio Molitor larva. The experimental design was a randomized block, where four different diets were tested for the development of T. molitor larvae. The control treatment consisted of wheat, soy, and corn, while the other three treatments consisted of diets supplemented with 50% of by-products derived from macauba, buriti, and cocoa extraction. Four treatments were carried out with four repetitions between May and June 2019. The data were analyzed using the ANOVA and Holm-Sidak tests, where p values <0.05 were considered statistically significant. T. molitor larvae completed their life cycle in all tested diets; however, the development in number of larvae in the diet supplemented with cocoa was reduced compared to the other groups. The flour of animals fed with a cocoa-based diet showed had a lower protein level when compared to the control group (p<0.001). Diets based on buriti, cacoa, and macaúba presented higher amounts of ether extract (p<0.018, <0.012, and <0.003, respectively). It is concluded that the supplementation of Tenebrios with diets based on buriti and macaúba can be a good strategy to improve the nutritional profile of T. molitor flour, providing favorable amounts of proteins and minerals. Regarding the use of cocoa, further studies to assess the presence of alkaloids and their direct effects on animal supplementation must be conducted, which may represent a limitation of this study.

Litomosoides brasiliensis (Nematoda: Onchocercidae) infecting chiropterans in the Legal Amazon region, Brazil / Litomosoides brasiliensis (Nematoda: Onchocercidae) em morcegos na Amazônia Legal, Brasil

Chiropterans play an important role in the maintenance of the environmental balance, since they are pollinators, seed dispersers and predators. They contribute to transmission and spreading of microorganisms such as helminths, fungi, protozoa, bacteria and virus. The aim of the present study was to investigate natural filariid infection among bats in the Legal Amazon region, Brazil, by means of parasitological and molecular analyses. Blood samples were collected from 82 bats for blood smears and for DNA extraction via the polymerase chain reaction (PCR) assay. Microfilariae were observed in blood smears from Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) and Glossophaga soricina (1). Five positive samples were detected through the PCR assay and four of these were also positive in blood smears. From genome sequencing and comparative analysis with sequences deposited in GenBank, one sample showed 99.31% similarity to the species Litomosoides brasiliensis. The present study expands the geographical distribution of L. brasiliensis, to include the state of Maranhão as an area of occurrence of this species and includes D. gnoma and A. fimbriatus as hosts in Brazil.(AU)

Os quirópteros desempenham um papel importante na manutenção do equilíbrio ambiental, pois são polinizadores, dispersores de sementes e predadores. Contribuem para a transmissão e disseminação de microrganismos, como helmintos, fungos, protozoários, bactérias e vírus. O objetivo do presente estudo foi investigar a infecção natural por filariídeos em morcegos na região da Amazônia Legal, Brasil, por meio de análises parasitológicas e moleculares. Amostras de sangue foram coletadas de 82 morcegos para confecção de esfregaços e para extração de DNA para a Reação em Cadeia da Polimerase (PCR). Microfilárias foram observadas em esfregaços sanguíneos de Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) e Glossophaga soricina (1). Cinco amostras positivas foram detectadas por meio do ensaio de PCR, e quatro destas também foram positivas em esfregaços de sangue. A partir do sequenciamento do genoma e análise comparativa com sequências depositadas no GenBank, uma amostra apresentou 99,31% de similaridade com a espécie Litomosoides brasiliensis. O presente estudo expande a distribuição geográfica de L. brasiliensis, abrangendo o estado do Maranhão como área de ocorrência desta espécie, e inclui D. gnoma e A. fimbriatus como hospedeiros no Brasil.(AU)

Biotecnologias na preservação de espermatozoides felinos / Biotechnologies in the preservation of feline sperm

O gato doméstico é a única espécie da família Felídea sem risco ou iminência de extinção, diferente da maior parte dos felinos selvagens. Desta forma, o desenvolvimento e aprimoramento de diferentes biotécnicas reprodutivas, são essenciais para a manutenção da qualidade reprodutiva, tendo em vista a preservação de espécies mais vulneráveis. Além disso, as biotécnicas do sêmen são para as tecnologias reprodutivas, como a inseminação artificial (IA) e a fertilização in vitro (FIV). Sendo assim, o objetivo deste compilado bibliográfico foi abordar as principais técnicas de colheita, análise e preservação de sêmen/espermatozoides felino, assim como o uso dessas células em IA e FIV. Para a colheita do sêmen felino, diferentes métodos têm sido aplicados: ejaculação farmacológica, eletroejaculação e vagina artificial. Em caso de óbito do reprodutor, os espermatozoides recuperados do epidídimo também apresentam viabilidade reprodutiva. Ademais, a cinética espermática avaliada pelo sistema CASA, a morfologia e a morfometria são as principais análises que demonstram a qualidade espermática e refletem na fertilidade do ejaculado. O sistema CASA também avalia a trajetória individual de cada espermatozoide, que ao se agrupar em clusters, demonstra a heterogeneidade do ejaculado nas subpopulações. Contudo, os diluentes para a conservação e refrigeração dos espermatozoides felinos e as curvas de congelação ainda não estão totalmente estabelecidos e influenciam diretamente a viabilidade dos espermatozoides criopreservados. Diante disso, os resultados da utilização do sêmen felino após criopreservação são inconsistentes, sendo necessários mais estudos para elucidar melhores curvas de congelação e meios de diluentes para viabilizar a preservação do material genético dos gatos.

The domestic cat is the only species of the Felidea family without risk or imminence of extinction, unlike most wild cats. Therefore, the development and improvement of different reproductive biotechnologies are essential for the maintenance of reproductive quality for the preservation of the most vulnerable species. Furthermore, semen biotechnologies are the basis for reproductive technologies such as artificial insemination (AI) and in vitro fertilization (IVF). Thus, the objective of this bibliographic compilation was to approach the main techniques of collection, analysis, and preservation of feline semen/sperm, as well as the use of these cells in AI and IVF. For feline semen collection, different methods have been applied: pharmacological ejaculation, electroejaculation, and artificial vagina. In case of death of the sire, sperm recovered from the epididymis also show reproductive viability. Moreover, the sperm kinetics evaluated by the CASA system, the morphology, and the morphometry are the main analyzes that demonstrate sperm quality and reflect on ejaculate fertility. The CASA system also evaluates the individual path of each sperm, which, when grouped into clusters, demonstrates the heterogeneity of the ejaculate in the subpopulations. However, diluents for the conservation and refrigeration of feline sperm and freezing curves are not yet fully established and directly influence the viability of cryopreserved sperm. Therefore, the results of using feline semen after cryopreservation are inconsistent, and further studies are needed to elucidate better freezing curves and diluents to enable the preservation of the genetic material of cats.

Quality indicators in colonoscopy: observational study in a supplementary health system

Purpose: Colorectal cancer is responsible for 9.4% of cancer deaths, and low polyp detection rate and cecal intubation rate increase the risks of interval colorectal cancer. Despite several population studies that address colonoscopy quality measures, there is still a shortage of these studies in Latin America. The aim of this study was to assess quality indicators in colonoscopy, enabling future strategies to improve colorectal cancer prevention. Methods: An observational retrospective study, in which all colonoscopies performed in 11 hospitals were evaluated through a review of medical records. Information such as procedure indication, colorectal polyp detection rate, cecal intubation rate, quality of colonic preparation, and immediate adverse events were collected and analyzed. Results: In 17,448 colonoscopies performed by 86 endoscopists, 57.9% were in patients aged 50 to 74 years old. Colon preparation was adequate in 94.4% procedures, with rates of cecal intubation and polyp detection of 94 and 36.6%, respectively. Acute adverse events occurred in 0.2%. In 53.9%, high-definition imaging equipment was used. The procedure location, colon preparation and high-definition equipment influenced polyp detection rates (p < 0.001). Conclusions: The extraction and analysis of electronic medical records showed that there are opportunities for improvement in colonoscopy quality indicators in the participating hospitals.

Avanços na andrologia de animais selvagens / Advances in wild animal andrology

As tecnologias de reprodução assistida (TRA) são de fundamental importância para a conexão de indivíduos em diferentes localidades, facilitando assim o intercâmbio genético e favorecendo a variabilidade genética de uma espécie. Por esta razão, as TRAS podem ser ferramentas importantes para a conservação de espécies ameaçadas de extinção. Apesar dos esforços nas últimas décadas, o avanço no desenvolvimento de tais tecnologias está aquém à urgência de reverter processos de baixa variabilidade genética em algumas espécies. A necessidade de refinamento das técnicas para as particularidades fisiológicas e comportamentais de cada espécie, somada à raridade de acesso aos animais são os principais fatores relacionados as dificuldades em se avançar com as TRAS. As técnicas mais recentes desenvolvidas para a recuperação de espermatozoides em animais selvagens são a colheita farmacológica, com uso de alfa-2-agonistas e a criopreservação / vitrificação testicular com posterior cultivo. Pouco de avançou, no entanto, em relação aos métodos de criopreservação, prevalecendo associação clássica de TRIS-gema-glicerol. Discutimos, então os métodos usados para acesso ao gameta masculino em espécies selvagens e suas aplicações na conservação animal.(AU)

Assisted reproduction technologies (ART) are of fundamental importance for connecting individuals in different locations, thus facilitating genetic exchange and favoring the genetic variability of a species. For this reason, TRAS can be important tools for the conservation of endangered species. Despite efforts in recent decades, the advance in the development of such technologies is short of the urgency of reversing processes of low genetic variability in some species. The need to refine the techniques for the physiological and behavioral particularities of each species, added to the rarity of access to animals, are the main factors related to the difficulties in advancing with TRAS. The most recent techniques developed for sperm collection in wild animals are pharmacological collection, with the use of alpha-2-agonists and testicular cryopreservation / vitrification with subsequent cultivation. Little progress has been made, however, in relation to cryopreservation methods, prevailing the classic association of TRIS-yolk-glycerol. We therefore discuss the methods used to access the male gamete in wild species and their applications in animal conservation.(AU)

DNA methylation level of gene SIRT1 in ram spermatozoa and relationship with fertilizing ability according to breed and age

Background: Effect of the epigenetic factors on the male fertility is well proofed. Sperm acts as a carrier of genetic material, and its DNA methylome can affect maternal pregnancy rate and offspring phenotype. However, the research on the DNA methylation in the spermatozoids of livestock males, in particular rams, is still limited. To best of our knowledge the data about as a global as well as gene specific DNA methylation in ram spermatozoa from different breeds and ages are missed in the scientific literature. The present study was designed to analyze the relationship between methylation levels of the important for spermatogenesis gene SIRT1 in spermatozoa and fertilizing ability of sperm in rams from different breeds and ages. Materials, Methods & Results: The ejaculates of 16 rams from Lacaune, East Friesian and Assaf breeds at age between 18 to 96 months were evaluated. The kinematic parameters of 2 semen samples from each animal were estimated by CASA. The separated spermatozoa were used for DNA extraction followed by bisulfite conversion. The DNA methylation of SIRT1 was detected through quantitative methylation-specific PCR using 2 sets of primers designed specifically for bisulfite-converted DNA sequences to attach methylated and unmethylated sites. The breed and age effect on the gene SIRT1 methylation by ANOVA was estimated. Experimental females included 393 clinically healthy milk ewes (Lacaune, n = 131; East Friesian sheep, n = 100 and Assaf, n = 162) in breeding season. Reproductive performances (conception rate at lambing, lambing percentage and fecundity) of ewes, inseminated by sperm of the investigated rams, were statistically processed. ANOVA showed that the animal breed influences significantly on the level of DNA methylation of gene SIRT1 in ram spermatozoa (P = 0.002) An average value of DNA methylation of SIRT1 in ram sperm from Lacaune breed was significantly higher than in Assaf and East Friesian (81.21 ± 15.1% vs 36.7 ± 14.2% and 38.3 ± 18.6 respectively, P < 0.01). The highest percent of SIRT1 methylation was observed in old animals compared to the young and middle-age. Moderate and strong correlations (r from 0.44 to 0.71, P < 0.05) between the methylation level of the SIRT1 gene in rams' sperm and reproductive parameters of inseminated ewes in all breeds were established. Discussion: Our data are the first message about the effect of breed on the specificity of DNA methylation of gene SIRT1 in ram spermatozoa. These results demonstrated an existence of the sheep breeds with high and low level of DNA methylation of gene SIRT1 in ram sperm. Although the effect of age on the methylation level in sperm is still discussable, our results showed a moderate correlation between age and methylation level of SIRT1 in spermatozoa of rams. Taking into account that DNA methylation in sperm is stabilized with puberty onset and is a heritable epigenetic modification, it can be a promising marker of sperm quality in animal breeding. In all investigated breeds the rams with relatively high level of DNA methylation of gene SIRT1 in spermatozoa (50-68%) demonstrated a high conception rate at lambing (> 70%). In conclusion, the DNA methylation level of the SIRT1 gene in ram spermatozoa is determined by both the breed and the age of the animals and correlates with fertilizing ability of sperm.

Molecular classification and comparative phylogeographic study of insectivorous bat species (Pipisitrellus coromandra) from Punjab, Pakistan / Classificação molecular e estudo filogeográfico comparativo de espécies de morcegos insetívoros (Pipistrellus coromandra) de Punjab, Paquistão

Molecular based identification of bat fauna in Pakistan has been relatively less explored. The current study was therefore planned to report for the first time the molecular classification of insectivorous bats (Pipistrellus coromandra) based on mitochondrion gene (COI) from Punjab, Pakistan. Specimens were collected from five different locations followed by DNA extraction with subsequent gene amplification and sequencing. All samples in the study had shown close identity matches with species (Pipistrellus coromandra) from India and (Pipistrellus tenuis) from Vietnam with percentage identity score of 96.11 and 95.58 respectively except one sequence which only revealed 86.78% identity match on Basic Local Alignment Search Tool (BLAST) and could only be assigned to genus level Pipistrellus sp. The results indicated negligible intra-population genetic distance among collected samples whereas the comparison with species from other countries had shown high intraspecific (P. coromandra) and interspecific (P. tenuis) mean genetic distances. The current study hence successfully proved the efficiency of COI gene as a molecular marker for species identification and in analyzing the patterns of genetic variation with species from other countries.

A identificação com base molecular da fauna de morcegos no Paquistão tem sido relativamente menos explorada. Portanto, o estudo atual foi planejado para relatar pela primeira vez a classificação molecular de morcegos insetívoros (Pipistrellus coromandra) com base no gene da mitocôndria (COI) de Punjab, Paquistão. As amostras foram coletadas em cinco locais diferentes, seguidas pela extração de DNA com subsequente amplificação e sequenciamento do gene. Todas as amostras no estudo mostraram coincidências de identidade próximas com espécies (Pipistrellus coromandra) da Índia e (Pipistrellus tenuis) do Vietnã, com pontuação de identidade percentual de 96,11 e 95,58, respectivamente, exceto uma sequência que revelou apenas 86,78% de correspondência de identidade na Ferramenta de Pesquisa de Alinhamento Local Básico (BLAST), a qual só poderia ser atribuída ao nível de gênero Pipistrellus sp. Os resultados indicaram distância genética intrapopulacional desprezível entre as amostras coletadas, enquanto a comparação com espécies de outros países mostrou altas distâncias genéticas intraespecíficas (P. coromandra) e interespecíficas (P. tenuis) médias. O presente estudo, portanto, comprovou com sucesso a eficiência do gene COI como marcador molecular para identificação de espécies e análise dos padrões de variação genética com espécies de outros países.

Isolation and amplification of Anaplasma marginale in chicken embryo fibroblast cell culture / Isolamento e amplificação de Anaplasma marginale em cultivos celulares de fibroblastos de embriões de galinhas

Anaplasma marginale (A. marginale) is a worldwide pathogen that infects a variety of ruminants, but mostly cattle. The present study aimed to describe an isolation technique for A. marginale, using chicken embryo Þ broblast (CEF) cell culture. Blood and tick samples were collected from 5 calves from 2 to 3 months old, which were considered to be infected with A.marginale due to anemia, jaundiced mucous membranes, and prostration. DNA extraction and PCR were performed for diagnosis using blood and tick samples. All tick and blood samples tested positive in PCR. Additionally, ticks were crushed with the aid of a blender for inoculation in CEF cell culture. After inoculation, the cultures were kept at 37ºC and 5% CO2 for 15 days. The cell supernatant of cell cultures was again analyzed using PCR and Wright stain method to conÞ rm A. marginale isolation. Cell cultures tested positive in PCR, and the presence of the agent was demonstrated by Wright stain. Therefore, by using CEF cell culture it was possible to isolate and amplify the A. marginale in a concentration of 1.3 x 107.2 bodies per mL. The CEF cells are undemanding and easy to preserve; they are an option for isolation and production of A. marginale under laboratory conditions.(AU)

Anaplasma marginale (A. marginale) é um patógeno mundial que infecta uma variedade de ruminantes, mas principalmente bovinos. O presente estudo teve como objetivo descrever uma técnica de isolamento para A. marginale, utilizando cultivo celular de Þ broblastos de embriões (CFE) de galinhas. Para isso, foram coletadas amostras de sangue e de carrapatos de 5 bezerros, entre 2 e 3 meses de idade, os quais, devido a anemia, icterícia de mucosas e prostração, foram considerados supostamente infectados com A. marginale. Ethics Approval This study was approved by Credenciamento Institucional para Atividades com Animais em Ensino ou Pesquisa (CIAEP: 02.0420.2021). Consent to participate Not applicable Consent to publish Not applicable Data availability Not applicable Para o diagnóstico, realizaram-se extração de DNA e posterior PCR a partir das amostras de sangue e de carrapatos coletados. Todos os carrapatos e amostras de sangue foram positivas para o teste de PCR. Além disso, os carrapatos foram triturados com o auxílio de um liquidiÞ cador para inoculação em CFE. Após a inoculação, as culturas foram mantidas a 37ºC e a 5% de CO2 durante 15 dias. O sobrenadante celular das culturas foi novamente analisado por PCR e pela técnica de coloração de Wright para conÞ rmar o isolamento de Anaplasma marginale. As culturas celulares foram po sitivas por PCR, e a presença do agente foi comprovada por meio da coloração de Wright. Portanto, utilizando CFE, foi possível isolar e ampliÞ car o A. marginale em uma concentração de 1,3x107,2 bactérias por ml. As células da CEF são pouco exigentes, de fácil manutenção e uma boa opção para isolamento e produção de A. marginale em condição laboratorial.(AU)