Interaction between methanotrophy and gastrointestinal nematodes infection on the rumen microbiome of lambs.

Complex crosstalk occurs between gastrointestinal nematodes and gut symbiotic microbiota, with consequences for animal metabolism. To investigate the connection between methane production and endoparasites, this study evaluated the effect of mixed infection with Haemonchus contortus and Trichostrongylus colubriformis on methanogenic and methanotrophic community in rumen microbiota of lambs using shotgun metagenomic and Real-time quantitative PCR (qPCR). The rumen content was collected from 6 Santa Inês lambs, (7 months old) before and after 42 days infection by esophageal tube. The metagenomic analysis showed that the infection affected the microbial community structure leading to decreased abundance of methanotrophs bacteria, i.e. α-proteobacteria and ß-proteobacteria, anaerobic methanotrophic archaea (ANME), protozoa, sulfate-reducing bacteria, syntrophic bacteria with methanogens, geobacter, and genes related to pyruvate, fatty acid, nitrogen, and sulfur metabolisms, ribulose monophosphate cycle and Entner-Doudoroff Pathway. Additionally, the abundance of methanogenic archaea and the mcrA gene did not change. The co-occurrence networks enabled us to identify the interactions between each taxon in microbial communities and to determine the reshaping of rumen microbiome associations by gastrointestinal nematode infection. Besides, the correlation between ANMEs was lower in the animal's post-infection. Our findings suggest that gastrointestinal parasites potentially lead to decreased methanotrophic metabolism-related microorganisms and genes.

Taxonomy and Functional Diversity in the Fecal Microbiome of Beef Cattle Reared in Brazilian Traditional and Semi-Intensive Production Systems.

The importance of beef production for economy of Brazil and the growing demand for animal protein across the globe warrant an improvement in the beef production system. Although most attention has been on modulation of the rumen microbiome to improve ruminant production, the role of the lower gut microbiome in host health and nutrition remains relatively unexplored. This work aimed to investigate the taxonomy and functional variations in the fecal microbiome of Brazilian beef cattle reared in two different production systems using a metagenomic approach. Sixty male beef cattle from six farms representing semi-intensive (I, n = 2) and traditional (T, n = 4) Brazilian beef production systems were enrolled in the study. Shotgun sequencing was used to characterize taxonomic and functional composition and diversity of the microbiome in fecal samples collected from each animal. Fecal samples were analyzed for copper (Cu), lead (Pb), nitrogen (N), phosphorous (P), selenium (Se), and zinc (Zn) and stable isotopes of carbon (13C) and nitrogen (15N). The fecal microbiome was influenced by the beef production systems with greater functional and lower taxonomic diversity in beef cattle feces from I systems compared with that from T systems. The concentration of N, P, and Zn was higher in beef cattle feces from I systems compared with that from T systems and was associated with taxonomic and functional profile of fecal microbiome in I system, suggesting the role of fecal nutrients in shaping system-specific microbiome. Semi-intensive management practices led to a more complex but less connected fecal microbiome in beef cattle. The microbial community in beef cattle feces from I systems was characterized by greater abundance of beneficial bacteria (phylum Firmicutes and butyrate-producing bacteria family Lachnospiraceae and genera Anaerostipes, Blautia, Butyrivibrio, Eubacterium, Roseburia, and Ruminococcus). In addition, the fecal abundance of microbial genes related to immune system, nutrient metabolism, and energy production was greater in beef cattle raised under I systems compared with that under T systems. Findings of the current study suggest that semi-intensive management practices could facilitate the development of a healthier and more efficient fecal microbiome in beef cattle by driving an increase in the abundance of beneficial bacteria and functional genes.

The effect of Haemonchus contortus and Trichostrongylus colubriforms infection on the ruminal microbiome of lambs.

We evaluated Haemonchus contortus (HC) and Trichostrongylus colubriformis (TC) infection on the ruminal microbial community of Santa Ines lambs to better understand the pathophysiology of parasite infections and the interactions among gastrointestinal nematodes and gut resident microbiota. In this study, 18 six months of age lambs were maintained for 34 days in individual pens divided into three treatments that included animals infected with HC and TC, and control (infection-free). Haematological, ruminal parameter and microbial nitrogen absorbed by pune derivatives, as well as enteric methane emission (CH4), were analysed, and the rumen microbial taxonomic and functional profile assessed by shotgun metagenomics. The analysis showed that total protein, albumin, urea, and butyrate level were lower in animals infected by both parasites, while HC infection also decreased the haemoglobin level. Both infected groups (TC and HC) increased the enteric methane emission (CH4). TC and HC infections increased the diversity and richness of functional microbial genes. Most alterations in the rumen microbiome composition of infected groups are associated with the suppression of microbes involved in microbial homeostasis maintenance and expansion of the archaeal community in the infected animals. Infection led to an increased abundance of nitrogen, amino acid, protein, and energy metabolism genes. Overall, TC and HC infection increased the enteric methane emission, negatively affected taxon's responsible for maintenance de rumen homeostasis and modulated some important genes related to protein and energy metabolism.

Cottonseed (gossypol) intake during gestation and lactation does affect the ovarian population in ewes and lambs?

The aim of this study was to evaluate if the cottonseed intake during gestation and lactation affects the ovarian population in ewes and lambs. Therefore, 39 ewes were evaluated during 10 months under two treatments: Cottonseed and soybeans. The quantification of ovarian follicular dynamics was analyzed by ultrasound and the determination of progesterone and estradiol levels was interpreted by radioimmunoassay. After weaning, ovaries of lambs (n = 10) were collected by ovariectomy and fixed for the assessment of follicular parameters as normality, classification, diameter, ultrastructure, stereology, and as well as immunoexpression of the α-estradiol receptor α (ER-α). The results showed that the cottonseed consumption altered neither the ovarian nor the hormonal follicular dynamics of Santa Inês ewes after calving and did not affect the normality, classification, diameter, stereology and follicular ultrastructure of offspring. Nevertheless, the offspring of ewes fed with cottonseed showed high ER-α immunoexpression in the ovarian structures. It is concluded that cottonseed did not affect the maternal-descendant follicular dynamics. However, lambs' ovaries had highest α-ER immunoexpression in oocytes, granulosa and theca cells and corpus luteum. This fact warns of a possible change in the future steroidogenic response of these lambs that had progenitors consuming cottonseed in their reproductive period.

Tannin supplementation modulates the composition and function of ruminal microbiome in lambs infected with gastrointestinal nematodes.

This study was carried out to evaluate the effects of tannin supplementation on ruminal microbiota of sixteen lambs infected and non-infected with Haemonchus contortus and Trichostrongylus colubriformis. Animals were fed with hay, concentrate and supplemented with Acacia mearnsii (A. mearnsii). The animals were divided into four treatments: two control groups without infection, either receiving A. mearnsii (C+) or not (C-), and two infected groups, one with A. mearnsii (I+) and another without A. mearnsii (I-). Ruminal short-chain fatty acids (SCFA) and metagenome sequencing of ruminal microbiota were used to evaluate the effect of tannin and infection on ruminal microbiome. For SCFA, differences were observed only with A. mearnsii. Total SCFA and acetate molar percentage were decreased in C+ and I+ (P<0.05). Butyrate, valerate and isovalerate were higher in lambs that received A. mearnsii in the diet (P<0.05). The infection changed the microbiome structure and decreased the abundance of butyrate-producing microorganisms. In addition, A. mearnsii supplementation also affected the structure the microbial community, increasing the diversity and abundance of the butyrate-producing and probiotics bacteria, amino acid metabolic pathways, purine, pyrimidine and sphingolipid metabolism. Together, our findings indicate that A. mearnsii supplementation modulates important groups related to nitrogen, amino acid, purine and pyrimidine metabolism, in rumen microbiome, affected by gastrointestinal nematodes infection in lambs.

Effects of Maternal Dietary Cottonseed on the Profile of Minerals in the Testes of the Lamb.

The use of cotton co-products in animal feed is restricted by the presence of gossypol, which is a toxic and highly reactive molecule of complex minerals. In mammals, part of the offspring phenotype is influenced by dam nutrition. The aim of this study was to investigate the effect of ewe diet, with and without cottonseed (gossypol), on the testicular development of lambs from birth to weaning through the assessment of lamb live weight (LW), macro and histological morphology of testes, and mass fraction of chemical elements by neutron activation analysis (NAA) and microprobe X-ray fluorescence spectroscopy, as well as the multielement distribution map in the testes. Eighteen lambs were used with an average LW at birth of 4 ± 1.0 kg. All lambs were offspring of Santa Inês ewes, fed on ration either with or without cottonseed during mating, gestation, and lactation, thus forming two treatments: control group (C) without cottonseed and treatment group with cottonseed (G). The animals were weighed from birth to 60 days of age, at fortnightly intervals. At 60 days of age, the lambs were orchidectomized to collect their testes for macroscopy, histological, neutron activation, and X-ray fluorescence analysis. Besides dry matter (DM), protein, ether extract (EE), calcium (Ca), and potassium (K) were higher in the ewe milk from the C group compared with the G group (P < 0.05). Lambs from the C group showed higher LW from 45 days onwards, as well as higher average daily gain when compared with the G group (P < 0.05). They also presented higher testicular weight, volume, length, width, as well as tubule and lumen diameters compared with lambs from the G treatment (P < 0.05). Multielementary NAA revealed higher mass fractions of rubidium (Rb), selenium (Se), and cesium (Cs) in the testes of lambs from the C group when compared with the G group, while G showed higher zinc (Zn) content (P < 0.05). No differences between treatments were found for element levels and distribution using X-ray fluorescence microanalysis (P > 0.05). In conclusion, the maternal cottonseed diet compromised performance and testes development of the lambs and reduce the content of Se in the testes.

Fatty acid profile and carcass characteristics in castrated and uncastrated hair lambs.

The aim of this study was to investigate the effects of castration on carcass characteristics and fatty acid profile of visceral fat and meat from lambs. Eighteen six-month-old Santa Inês male lambs (18.9 ± 2.4 kg of body weight (BW)) were used. Animals were assigned to two treatments according to their sexual condition: uncastrated (U) or castrated (C). During a 98-day experimental period, animals were kept on an Andropogon gayanus grass pasture area of 1 ha and supplemented with 200 g/animal/day of concentrate mixture. Water and mineral salt were available ad libitum. The lambs were weighed fortnightly, and at the last day of the trial, they were slaughtered for evaluation of carcass characteristics and fatty acids profile of perirenal fat and longissimus lumborum muscle samples. Castrated lambs showed higher BW than U during most part of the experiment (p < 0.05). Fat deposition was higher in C lambs as evidenced by their increased carcass fat cover. Meat from U lambs showed lower content of C16:0 and higher polyunsaturated fatty acids (PUFA) (p < 0.05) than that from C (U, 14.3 and C, 10.5%). Conjugated linoleic acid (CLA) content was not affected by castration (p > 0.05) (U, 0.74 and C, 0.76%). The cis-9, cis-12 C18:2n-6 (U, 10.6 and C, 6.86%) fatty acid and the PUFA:SFA (saturated fatty acids) ratio (U, 0.36 and C, 0.25) were higher in the muscle of U lambs (p < 0.05), indicating that the meat from U animals may provide more benefits to human health than that of C.

Morphological-metric, ultrastructural and immunohistochemical effects of gossypol on cultured granulosa cells and oocytes of ewes using MOEPF.

Manipulation of oocytes enclosed in preantral follicles (MOEPF) allows for analyzing follicular development and use of this biotechnology in the pre-analysis of the beneficial or toxic effects of bio-products on granulosa cells and oocytes at different developmental stages. In this study, there was evaluation of the effects of gossypol by culturing granulosa cells and oocytes in ewe ovarian tissues. Ovarian tissues were cultured with gossypol at 37 °C, in humidified air and 5% CO2. Variables that were evaluated were morphology, morphometry, ultrastructure and abundance of estradiol receptor α (α-ER). There were no differences in developmental characteristics when there was treatment with any of the gossypol doses that were evaluated. Immunostaining indicated that when the gossypol dose increases, the abundance of α-ER also increases in the cytoplasm, nucleus, and granulosa cells. Findings with the ultrastructural analysis indicated that for granulosa cells there was fewer cells and greater disorganization and a lack of structural integrity of follicular cell layers as a result of all gossypol treatments. The culture of oocytes in preantral ovarian follicles in presence of gossypol did not affect the morphological-metric structure at the doses evaluated. The findings with evaluation of ultrastructural and immunohistochemical structures indicated granulosa cells and α-ER were affected by the treatments with gossypol indicating there were effects of this compound on ovarian function in sheep. This study indicated there is a toxic action of gossypol when using the biotechnology, MOEPF. Thus, gossypol negatively affects granulosa cell development and structural integrity of preantral follicles in sheep.

Use of multivariate analyses for determining heat tolerance in Brazilian cattle.

Adaptability can be evaluated by the ability of an animal to adjust to environmental conditions and is especially important in extreme weather conditions such as that found in tropical Brazil. A multivariate analysis using physical and physiological traits in exotic (Nellore and Holstein) and naturalized (Junqueira, Curraleira, Mocho Nacional, Crioula Lageana, and Pantaneira) cattle breeds was carried out in the Federal District of Brazil to test and determine which traits are important in the adaptation of animal to heat stress as well as the ability of these traits and statistical techniques to separate the breeds studied. Both physical and physiological traits were measured on three occasions and included body measurements, skin and hair thickness, hair number and length, pigmentation, sweat gland area as well as heart and breathing rates, rectal temperature, sweating rate, and blood parameters. The data underwent multivariate statistical analyses, including cluster, discriminate, and canonical procedures. The tree diagram showed clear distances between the groups studied, and canonical analysis was able to separate individuals in groups. Coat traits explained little variation in physiological parameters. The traits which had higher discriminatory power included packed cell volume, shoulder height, mean corpuscular volume, body length, and heart girth. Morphological and physiological traits were able to discriminate between the breeds tested, with blood and size traits being the most important. More than 80% of animals of all breeds were correctly classified in their genetic group.

Use of heat tolerance traits in discriminating between groups of sheep in central Brazil.

The animal and its environment make up an integrated system, where each acts on the other. Tropical regions are characterized by high levels of solar radiation and environmental temperature which may adversely affect animal production. This study carries out a multivariate analysis of physical and physiological traits in sheep in the Federal District of Brazil to test the ability to separate groups of animals and determine which traits are most important in the adaptation of animal to heat stress. The variables studied included coat thickness, number and length of hairs, pigmentation of the skin and coat, number of sweat glands as well as heart and respiratory rates, rectal and skin temperatures, sweating rate, and blood parameters. Five groups of ten animals were used depending on breed (Bergamasca, crossbred, or Santa Inês) or coat color (Santa Inês--brown, white, and black). The data underwent multivariate statistical analyses including cluster, discriminate, and canonical, using Statistical Analysis System--SAS®. The tree diagram showed clear distances between groups studied and canonical analysis was able to separate individuals in groups, especially Bergamasca and white Santa Inês. The canonical correlation redundancy analysis showed that coat reflectance as well as hair length and number of hairs per unit area were the most useful in explaining changes in physiological traits. Skin and coat traits such as hair length, coat reflectance, percentage of epithelial area occupied by sweat glands, skin reflectance and thickness, as well as heart and breathing rates were the most important in separating these groups.