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1.
Arch Microbiol ; 202(7): 1861-1872, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32448959

RESUMO

In addition to a wide variety of anaerobic and facultative anaerobic bacteria, camel rumen also harbors a diverse of eukaryotic organisms. In the present study, the eukaryotic communities of camel rumen were characterized using 18S rRNA amplicon sequencing. Metagenomic DNA was isolated from rumen samples of fourteen adult Bikaneri and Kachchhi breeds of camel fed different diets containing Jowar, Bajra, Maize, and Guar. Illumina sequencing generated 27,161,904 number of reads corresponding to 1543 total operational taxonomic units (OTUs). Taxonomic classification of community metagenome sequences from all the samples revealed the presence of 92 genera belonging to 16 different divisions, out of which Ciliophora (73%), Fungi (13%) and Streptophyta (9%) were found to be the most dominant. Notably, the abundance of Ciliophora was significantly higher in the case of Guar feed, while Fungi was significantly higher in the case of Maize feed, indicating the influence of cellulose and hemicellulose content of feedstuff on the composition of eukaryotes. The results suggest that the camel rumen eukaryotes are highly dynamic and depend on the type of diet given to the animal. Pearson's correlation analysis suggested the ciliate protozoa and fungi were negatively correlated with each other. To the best of our knowledge, this is first systematic study to characterize camel rumen eukaryotes, which has provided newer information regarding eukaryotic diversity patterns amongst camel fed on different diets.


Assuntos
Camelus/microbiologia , Camelus/parasitologia , Cilióforos , Dieta , Fungos , Rúmen/microbiologia , Rúmen/parasitologia , Animais , Cilióforos/classificação , Cilióforos/genética , Fungos/classificação , Fungos/genética , Sequenciamento de Nucleotídeos em Larga Escala , Metagenoma , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
2.
PLoS One ; 15(4): e0231516, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32298315

RESUMO

The aim of this study was to evaluate the effects of medicinal herbal mixtures rich in phenolic, flavonoid and alkaloid compounds on ruminal fermentation and microbial populations, and fatty acid (FA) concentrations and lipid oxidation in tissues of lambs infected with the gastrointestinal nematode (GIN) parasite (Haemonchus contortus). Parallel in vitro and in vivo studies were performed using two different herbal mixtures (Mix1 and Mix2). The in vitro study was conducted in a 2 (infection status; non-infected versus infected) × 3 (diets; control, Mix1 and Mix2) factorial design. In the in vivo study, 24 lambs were equally divided into four treatments: non-infected lambs fed a control diet, infected lambs fed the control diet, infected lambs fed a diet with Mix1 and infected lambs fed a diet with Mix2. Herbal mixtures (100 g dry matter (DM)/d) were added to the basal diets of meadow hay (ad libitum) and a commercial concentrate (500 g DM/d). The experimental period lasted for 70 days. Ruminal fermentation characteristics and methane production were not affected by infection in vivo or in vitro. Both herbal mixture supplementation increased total volatile fatty acid (VFA) concentrations (P < 0.01) and DM digestibility (P < 0.01) in vitro. Archaea population was slightly diminished by both herbal mixtures (P < 0.05), but they did not lower methane production in vitro or in vivo (P > 0.05). Infection of H. contortus or herbal mixtures modulated FA proportion mainly in the liver, especially the long chain FA proportion. Concentrations of thiobarbituric acid reactive substances (TBARS) in serum were significantly higher after 70 days post-infection in the infected lambs. Herbal Mix1 supplementation reduced TBARS concentrations in meat after seven days of storage. In conclusion, supplementing of herbal mixtures to the diets of GIN parasite infected lambs did not affect the basic ruminal fermentation parameters. Herbal mixtures may improve few FA proportions mainly in liver as well as decrease lipid oxidation in meat.


Assuntos
Suplementos Nutricionais , Fermentação/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Hemoncose/veterinária , Haemonchus , Metabolismo dos Lipídeos/efeitos dos fármacos , Plantas Medicinais/metabolismo , Rúmen/metabolismo , Doenças dos Ovinos/parasitologia , Animais , Dieta/veterinária , Ácidos Graxos/metabolismo , Feminino , Hemoncose/metabolismo , Rúmen/microbiologia , Rúmen/parasitologia , Ovinos/metabolismo , Ovinos/microbiologia , Ovinos/parasitologia , Doenças dos Ovinos/metabolismo
3.
Mol Phylogenet Evol ; 143: 106687, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31740334

RESUMO

Rumen ciliates are a specialized group of ciliates exclusively found in the anaerobic, carbohydrate-rich rumen microenvironment. However, the molecular and mechanistic basis of the physiological and behavioral adaptation of ciliates to the rumen microenvironment is undefined. We used single-cell transcriptome sequencing to explore the adaptive evolution of three rumen ciliates: two entodiniomorphids, Entodinium furca and Diplodinium dentatum; and one vestibuliferid, Isotricha intestinalis. We found that all three species are members of monophyletic orders within the class Litostomatea, with E. furca and D. dentatum in Entodiniomorphida and I. intestinalis in Vestibuliferida. The two entodiniomorphids might use H2-producing mitochondria and the vestibuliferid might use anaerobic mitochondria to survive under strictly anaerobic conditions. Moreover, carbohydrate-active enzyme (CAZyme) genes were identified in all three species, including cellulases, hemicellulases, and pectinases. The evidence that all three species have acquired prokaryote-derived genes by horizontal gene transfer (HGT) to digest plant biomass includes a significant enrichment of gene ontology categories such as cell wall macromolecule catabolic process and carbohydrate catabolic process and the identification of genes in common between CAZyme and HGT groups. These findings suggest that HGT might be an important mechanism in the adaptive evolution of ciliates to the rumen microenvironment.


Assuntos
Cilióforos/genética , Rúmen/parasitologia , Transcriptoma , Adaptação Fisiológica , Anaerobiose , Animais , Metabolismo dos Carboidratos , Celulases/genética , Cilióforos/classificação , Cilióforos/fisiologia , Transferência Genética Horizontal , Glicosídeo Hidrolases/genética , Filogenia , Poligalacturonase/genética , RNA-Seq , Rúmen/metabolismo , Análise de Célula Única
4.
J Dairy Sci ; 103(3): 2373-2386, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31882219

RESUMO

Wheat is the most common concentrate fed to grazing dairy cows in Australia, but no studies have examined the effects of wheat proportion in a pasture-based diet on milk production and methane emissions. In this 47-d experiment, 32 Holstein dairy cows were offered 1 of 4 diets during d 1 to 36. Cows in each of the dietary treatment groups were individually offered no wheat (W0) or wheat at 3 kg of dry matter (DM)/d (W3), 6 kg of DM/d (W6), or 9 kg of DM/d (W9). The remainder of the diet was 2.2 kg of DM of concentrate mix and freshly harvested perennial ryegrass (Lolium perenne) such that all individual cows were offered a total diet of approximately 20.2 kg of DM/d. From d 37 to 47 the diets of cows receiving treatments W0 and W3 remained unchanged, but cows in treatments W6 and W9 received the W3 diet. Individual cow feed intakes, milk yields, milk compositions, and methane emissions were measured for d 31 to 35 (period 1) and d 45 to 47 (period 2). During period 1, the mean intakes of cows offered the W0, W3, W6, and W9 diets were 19.2, 20.4, 20.2, and 19.8 kg of DM/d. Diet caused differences in energy-corrected milk, and means for W0, W3, W6, and W9 were 29.5, 32.4, 33.0, and 32.9 kg/d, respectively. Milk fat percentage differed with respective means of 3.93, 3.94, 3.69, and 3.17. Diets also caused differences in methane emissions, with means for W0, W3, W6, and W9 of 440, 431, 414, and 319 g/d. During period 1, the cows fed the W9 diet produced less methane and had lower methane yields (g/kg of DMI) and intensities (g/kg of energy-corrected milk) than cows fed the W3 diet. However, in period 2 when the wheat intake of cows in the W9 treatment was reduced to the same level as in the W3 treatment, their methane emissions, yields, and intensities were similar to those offered the W3 treatment, yet protozoa numbers in ruminal fluid were still much lower than those in cows offered the W3 treatment. Our research shows that for diets based on perennial ryegrass and crushed wheat, only the diet containing more than 30% crushed wheat resulted in substantially depressed milk fat concentration and reduced methane emissions, methane yield, and methane intensity. Thus, although feeding a diet with a high proportion of wheat can cause substantial methane mitigation, it can come at the cost of depression in milk fat concentration.


Assuntos
Ração Animal , Bovinos , Dieta/veterinária , Metano/biossíntese , Leite , Rúmen/parasitologia , Triticum , Animais , Austrália , Cilióforos/isolamento & purificação , Indústria de Laticínios , Feminino , Herbivoria , Lactação , Lolium , Poaceae
5.
BMC Vet Res ; 15(1): 360, 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31640708

RESUMO

BACKGROUND: Paramphistomosis is a pathogenic disease of domesticated ruminants, causing great economic loss in dairy industry and meat production. It is considered as a neglected tropical disease with highest prevalence throughout tropical and subtropical regions, particularly in Africa, Asia, Europe, and Australia. There are few trematocidal drugs available in the market. Most are resistant and have elevated side effects. Therefore, alternative trematocidal drugs need to discover. This study was conducted to evaluate three plants leaf extracts (from Senna alata, S. alexandrina, and S. occidentalis) as effective remedies against gastrointestinal trematode parasite (Paramphistomum gracile) of ruminants. Live adult parasites were collected in 0.1 M phosphate-buffered saline (PBS) from fresh autopsied goat's rumen. Parasites were incubated in leaf extracts of S. alata, S. alexandrina and, S. occidentalis individually and in combination (1:1) ratio at 37 ± 1°C. Treatment media contain extracts at different concentrations (10, 20 and 40 mg/mL) in 10 mL of 0.1 M PBS with 1% dimethylsulphoxide (DMSO). Parasites in control group were incubated in PBS without extract. The efficacy of three Senna extracts was evaluated on the basis of dose-dependent motility and mortality of the trematode. Immediately after paralysis, all treated parasites were collected for histology, SEM and biochemical study. RESULTS: Dose-dependent efficacy was observed in terms of motility and time of mortality in all treated parasites after exposure in various concentrations of the Senna plant extracts. S. occidentalis and S. alexandrina showed better efficacy in combination than comparing with individual treatment groups. Histological study and scanning electron microscopic observations revealed conspicuous deformity of surface architecture in all treated parasites. Scanning electron micrographs also revealed shrinkage, vacuolization, infoldings and blebbings on the body surface of treated worms. Activities of tegumental enzymes were inhibited in all treatment groups compared to control. CONCLUSION: The overall findings from this study revealed that all three Senna leaf extracts individually and in combination showed potential antitrematocidal activity against Paramphistomum gracile by damaging body tegument and neural propagation. Thus, this study confirmed that all three Senna extracts can be considered as a potential drug-like candidate in indigenous system of traditional medicine against trematode infections in livestock.


Assuntos
Anti-Helmínticos/farmacologia , Paramphistomatidae/efeitos dos fármacos , Extratos Vegetais/farmacologia , Senna (Planta)/química , Animais , Anti-Helmínticos/isolamento & purificação , Etnofarmacologia , Cabras , Paramphistomatidae/isolamento & purificação , Paramphistomatidae/ultraestrutura , Extratos Vegetais/isolamento & purificação , Rúmen/parasitologia , Infecções por Trematódeos/tratamento farmacológico , Infecções por Trematódeos/veterinária
6.
Artigo em Alemão | MEDLINE | ID: mdl-31212344

RESUMO

Paramphistosis is a globally occurring parasitic disease in various ruminants caused by a range of rumen flukes (including Paramphistomum cervi, Calicophoron daubneyi and Paramphistomum leydeni). In Europe, local occurrences of rumen fluke infection in domestic and wild ruminants have been described for decades. There is now evidence that paramphistomidosis is gaining in importance, because high prevalence rates were reported in the United Kingdom, Ireland, France, Spain, Belgium and The Netherlands. Current prevalence data from Germany are lacking. In recent investigations in northern Germany, Hesse and Bavaria, C. daubneyi was detected, which is currently the most prevalent rumen fluke in Europe. The development of therumen fluke is linked to aquatic snails as intermediate hosts. C. daubneyi and the liver fluke Fasciola hepatica share in the course of their development the same intermediate snail host, Galba truncatula. The definitive ruminant host takes up infective metacercaria. In the small intestine, the young flukes excyst and attach to the duodenum. Subsequently, they migrate to the rumen, where, as adults, they begin to release eggs. The infection can lead to severe diarrhea during the intestinal phase and death at high infection intensity. Ruminal paramphistomidosis is subclinical in most cases. Currently, coproscopic detection by the sedimentation method is the available diagnostic tool. Because of similar morphology, there is a risk of confusion with the eggs of the liver fluke F. hepatica. Paramphistomidosis can be treated with oxyclozanide. There are conflicting results regarding the effectiveness of other drugs. Therefore, prophylaxis of this parasitosis is important. Because of the similar epidemiology, control recommendations are based on those for the prevention of fasciolosis. Whether paramphistomidosis is also an emerging infectious disease in Germany cannot be currently assessed.


Assuntos
Paramphistomatidae , Infecções por Trematódeos , Animais , Anti-Helmínticos/uso terapêutico , Alemanha , Rúmen/parasitologia , Rúmen/patologia , Ruminantes , Infecções por Trematódeos/diagnóstico , Infecções por Trematódeos/tratamento farmacológico , Infecções por Trematódeos/epidemiologia , Infecções por Trematódeos/veterinária
7.
J Appl Microbiol ; 127(3): 670-682, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31165532

RESUMO

AIMS: Experiments were designed to determine the effects of different chemical inhibitors of lysozyme and peptidases on rumen protozoa and the associated prokaryotes, and in vitro fermentation using Entodinium caudatum as a model protozoan species. METHODS AND RESULTS: Imidazole (a lysozyme inhibitor), phenylmethylsulphonyl fluoride (PMSF, a serine peptidase inhibitor) and iodoacetamide (IOD, a cysteine peptidase inhibitor) were evaluated in vitro both individually and in two- and three-way combinations using E. caudatum monocultures with respect to their ability to inhibit the protozoan and their effect on feed digestion, fermentation and the microbiota. All the three inhibitors, both individually and in combination, decreased E. caudatum counts (P < 0·001), and IOD and its combinations with the other inhibitors significantly (P < 0·01) decreased ammonia concentration, with the two- and three-way combinations showing additive effective. Feed digestion was not affected, but fermentation and microbial diversity were affected mostly by PMSF, IOD and their combinatorial treatments potentially due to the overgrowth of Streptococcus luteciae accompanying with the disappearance of host ciliates. CONCLUSIONS: Entodinium caudatum depends on lysozyme and peptidase for digestion and utilization of the engulfed microbes and specific inhibition of these enzymes can inhibition E. caudatum without adversely affecting feed digestion or fermentation even though they changed the microbiota composition in the cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: The peptidase inhibitors may have the potential to be used in controlling rumen protozoa to improve ruminal nitrogen utilization efficiency.


Assuntos
Cilióforos/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Muramidase/antagonistas & inibidores , Inibidores de Proteases/farmacologia , Amônia/metabolismo , Animais , Cilióforos/enzimologia , Cilióforos/crescimento & desenvolvimento , Cilióforos/microbiologia , Digestão/efeitos dos fármacos , Fermentação/efeitos dos fármacos , Imidazóis/farmacologia , Iodoacetamida/farmacologia , Microbiota/efeitos dos fármacos , Fluoreto de Fenilmetilsulfonil/farmacologia , Rúmen/parasitologia
8.
Vet Parasitol ; 268: 9-15, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30981312

RESUMO

The prevalence of C. daubneyi infection in the United Kingdom has increased, but despite the potential for rumen flukes to cause production loss in ruminant livestock, understanding of their emergence and spread is poor. Here we describe the development of a method to explore the multiplicity of C. daubneyi infection and patterns of the parasite's emergence and spread, based on Illumina MiSeq deep sequencing of meta barcoded amplicons of a fragment of the cytochrome c oxidase subunit I (mt-COX-1) locus. Our results show high levels of genetic diversity in 32 C. daubneyi populations derived from finished prime cattle consigned to slaughter from northern United Kingdom. The results are consistent with a single introduction of C. daubneyi infection to some of the farms where the cattle had been grazed during their lifetime and multiple introductions to most. The results illustrate the impact of high levels of animal movements in the United Kingdom, whereby multiple common mt-COX-1 haplotypes were identified in 26 populations in the absence of geographical clustering of clades. This has implications for the adaptability of environmental and intermediate host stages of the parasite to changing climatic and animal management conditions, or of parasitic stages to exposure to anthelmintic drugs; potentially allowing for greater pathogenicity, or the development of anthelmintic resistance, respectively.


Assuntos
Doenças dos Bovinos/epidemiologia , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Paramphistomatidae/genética , Rúmen/parasitologia , Infecções por Trematódeos/veterinária , Animais , Bovinos/parasitologia , Doenças dos Bovinos/parasitologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fezes/parasitologia , Haplótipos , Gado/parasitologia , Prevalência , Infecções por Trematódeos/epidemiologia , Reino Unido/epidemiologia
9.
J Dairy Sci ; 102(5): 4205-4208, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30879814

RESUMO

Glycogen is an α-glucan produced by rumen microbes from various feed carbohydrates. It may be digested ruminally or intestinally to provide nutrients. The physicochemical and enzymatic hydrolysis characteristics of microbial glycogen have not been described in detail, but do influence its conversion to absorbable nutrients in vivo, its nutritional comparability with plant starch sources, and its accurate analysis in vitro. The objectives of this study were to determine presence or absence of a gelatinization response and to describe enzymatic digestion characteristics of glycogen granules isolated from ruminal protozoa obtained from lactating dairy cows. Protozoal glycogen granules were determined to be 98.3% α-glucan. Granules displayed gelatinization, the breaking of hydrogen bonds between molecules or branches, at 65°C compared with purified wheat and corn starches, which initiated gelatinization at 50 and 65°C, respectively. Digestion of ungelatinized samples with amyloglucosidase for 2 h at 39°C showed approximately 3-fold greater hydrolysis to glucose for protozoal glycogen (25.2% of dry matter; DM) than for wheat (9.9% of DM) or corn (8.2% of DM) starches. Based on enzymatic digestion results, protozoal glycogen may be more readily digested than intact corn or wheat starches and should be gelatinized or the hydrogen bonds otherwise disrupted to allow more complete recovery in enzymatic analysis.


Assuntos
Bovinos/metabolismo , Cilióforos/enzimologia , Glicogênio/metabolismo , Rúmen/metabolismo , Ruminação Digestiva , Animais , Feminino , Hidrólise , Lactação , Rúmen/parasitologia , Amido/metabolismo , Triticum , Zea mays
10.
Sci Rep ; 9(1): 2440, 2019 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-30792418

RESUMO

Although the potential of plants extracts to improve feed efficiency and animal productivity and decrease methane emissions by enteric fermentation has been shown, the information available is often contradictory which has been attributed to differences in the complex mixture of bioactive compounds and their interactions. Understanding the degree to which structural features in a compound may affect the biological activity of an extract is essential. We hypothesised that relative small variations in the structure of a compound can have a significant influence on the ability of the derivatives to alter fermentation in the rumen. Nine compounds were synthetized from the natural alkaloid haemanthamine and tested in vitro for their effects on rumen protozoa and fermentation parameters. Our results showed that simple esterifications of haemanthamine or its derivative dihydrohaemanthamine with acetate, butyrate, pivalate or hexanoate led to compounds that differed in their effects on rumen fermentation.


Assuntos
Alcaloides de Amaryllidaceae/química , Alcaloides de Amaryllidaceae/farmacologia , Ração Animal , Fermentação/efeitos dos fármacos , Fenantridinas/química , Fenantridinas/farmacologia , Rúmen/efeitos dos fármacos , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Bovinos , Dieta , Técnicas In Vitro , Testes de Sensibilidade Parasitária , Plantas/química , Infecções Protozoárias em Animais/patologia , Infecções Protozoárias em Animais/prevenção & controle , Rúmen/metabolismo , Rúmen/microbiologia , Rúmen/parasitologia
11.
Parasitol Res ; 118(2): 707-710, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30607609

RESUMO

A crucial step in the infection process of grazing ruminants by gastro-intestinal nematodes is the exsheathment of the infective third-stage larva following ingestion. Recently, heat shock was shown to play an important role in the carbon dioxide (CO2)-dependent exsheathment response in Haemonchus contortus. The current in vitro study set out to evaluate the role of heat shock in other abomasal species. In rumen fluid, all species tested exsheathed rapidly and efficiently in response to heat shock and CO2. This response was significantly higher compared to slow temperature changes, supporting the hypothesis that heat shock plays an important role in vivo. However, in artificial buffer, the effect of heat shock was species-dependent. For H. contortus and Ostertagia leptospicularis, the response in artificial buffer was similar to rumen fluid. In contrast, Ostertagia ostertagi and Teladorsagia circumcincta exsheathment was significantly lower and/or slower in artificial buffer, and there was no benefit of heat shock. For these two species, it appears that there are co-factors in the rumen fluid, in addition to heat shock and CO2, contributing to exsheathment. Overall, the data indicate that there are significant differences between abomasal species in their response to exsheathment triggers.


Assuntos
Abomaso/parasitologia , Dióxido de Carbono/farmacologia , Haemonchus/fisiologia , Resposta ao Choque Térmico/fisiologia , Ostertagia/fisiologia , Trichostrongyloidea/fisiologia , Animais , Trato Gastrointestinal/parasitologia , Larva/fisiologia , Rúmen/parasitologia , Ruminantes/parasitologia
12.
BMC Microbiol ; 18(1): 212, 2018 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-30545295

RESUMO

BACKGROUND: Tibetan sheep (TS) and Gansu Alpine Finewool sheep (GS) are both important plateau sheep raised and fed on the harsh Qinghai-Tibetan Plateau, China. Rumen methanogen and protozoal communities of plateau sheep are affected by their hosts and living environments, and play important roles in ruminant nutrition and greenhouse gas production. However, the characteristics, differences, and associations of these communities remain largely uncharacterized. RESULTS: The rumen methanogen and protozoal communities of plateau sheep were investigated by 16S/18S rRNA gene clone libraries. The predominant methanogen order in both sheep species was Methanobacteriales followed by Methanomassiliicoccales, which is consistent with those seen in global ruminants. However, the most dominant species was Methanobrevibacter millerae rather than Methanobrevibacter gottschalkii seen in most ruminants. Compared with GS and other ruminants, TS have more exclusive operational taxonomic units and a lower proportion (64.5%) of Methanobrevibacter. The protozoa were divided into Entodiniomorphida and Vestibuliferida, including nine genera and 15 species. The proportion of holotrich protozoa was much lower (1.1%) in TS than ordinary sheep. The most predominant genus was Entodinium (70.0%) in TS and Enoploplastron (48.8%) in GS, while the most common species was Entodinium furca monolobum (43.9%) and Enoploplastron triloricatum (45.0%) in TS and GS, respectively; Entodinium longinucleatum (22.8%) was only observed in TS. LIBSHUFF analysis indicated that the methanogen communities of TS were significantly different from those of GS, but no significant differences were found in protozoal communities. CONCLUSION: Plateau sheep have coevolved with unique rumen methanogen and protozoal communities to adapt to harsh plateau environments. Moreover, the host appears to have a greater influence on rumen methanogen communities than on rumen protozoal communities. The observed associations of methanogens and protozoa, together with the findings of previous studies on methane emissions from ruminant livestock, revealed that the lower proportion of Methanobrevibacter and holotrich protozoa may be responsible for the lower methane emission of TS. These findings facilitate our understanding of the rumen microbial ecosystem in plateau sheep, and could help the development of new strategies to manipulate rumen microbes to improve productivity and reduce the emission of greenhouse gases.


Assuntos
Euryarchaeota/metabolismo , Metano/metabolismo , Methanobrevibacter/metabolismo , Rúmen/microbiologia , Rúmen/parasitologia , Trichostomatida/metabolismo , Animais , Euryarchaeota/classificação , Euryarchaeota/genética , Euryarchaeota/isolamento & purificação , Comportamento Alimentar , Methanobrevibacter/classificação , Methanobrevibacter/genética , Methanobrevibacter/isolamento & purificação , Filogenia , Ovinos/microbiologia , Ovinos/parasitologia , Ovinos/fisiologia , Tibet , Trichostomatida/genética , Trichostomatida/isolamento & purificação
13.
Parasit Vectors ; 11(1): 617, 2018 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-30509301

RESUMO

BACKGROUND: Diseases caused by parasitic flatworms of rumen tissues (paramphistomosis) are a significant threat to global food security as a cause of morbidity and mortality in ruminant livestock in subtropical and tropical climates. Calicophoron daubneyi is currently the only paramphistome species commonly infecting ruminant livestock in temperate European climates. However, recorded incidences of C. daubneyi infection in European livestock have been increasing over the last decade. Whilst clinical paramphistomosis caused by adult worms has not been confirmed in Europe, fatalities have been attributed to severe haemorrhagic enteritis of the small intestine resulting from the migration of immature paramphistomes. Large numbers of mature adults can reside in the rumen, yet to date, the impact on rumen fermentation, and consequently on productivity and economic management of infected livestock, have not been resolved. Limited publicly available nucleotide and protein sequences for C. daubneyi underpin this lack of biological and economic understanding. Here we present for the first time a de novo assembled transcriptome, with functional annotations, for adult C. daubneyi, which provides a reference database for protein and nucleotide sequence identification to facilitate fundamental biology, anthelmintic, vaccine and diagnostics discoveries. RESULTS: This dataset identifies a number of genes potentially unique to C. daubneyi and, by comparison to an existing transcriptome for the related Paramphistomum cervi, identifies novel genes which may be unique to the paramphistome group of platyhelminthes. Additionally, we present the first coverage of the excretory/secretory and soluble somatic proteome profiles for adult C. daubneyi and identify the release of extracellular vesicles from adult C. daubneyi parasites during in vitro, ex-host culture. Finally, we have performed the first analysis of rumen fluke impacting upon rumen fermentation parameters using an in vitro gas production study resulting in a significant increase in propionate production. CONCLUSIONS: The resulting data provide a discovery platform (transcriptome, proteomes, EV isolation pipeline and in vitro fermentation system) to further study C. daubneyi-host interaction. In addition, the acetate: propionate ratio has been demonstrated to decrease with rumen fluke infection suggesting that acidotic conditions in the rumen may occur.


Assuntos
Doenças dos Bovinos/parasitologia , Gado/parasitologia , Paramphistomatidae/genética , Paramphistomatidae/metabolismo , Rúmen/parasitologia , Infecções por Trematódeos/veterinária , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/metabolismo , Europa (Continente)/epidemiologia , Vesículas Extracelulares , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/metabolismo , Genes de Helmintos , Proteínas de Helminto , Incidência , Redes e Vias Metabólicas/genética , Proteômica , Rúmen/metabolismo , Transcriptoma , Infecções por Trematódeos/epidemiologia , Infecções por Trematódeos/parasitologia
14.
Parasitol Res ; 117(11): 3675-3678, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30264327

RESUMO

The first step in the infection process of grazing ruminants by gastrointestinal nematodes is the exsheathment of the third-stage larva (L3). Exsheathment of various species can be achieved in vitro using carbon dioxide (CO2) under the appropriate temperature and pH conditions. However, it remains unclear whether elevated CO2 levels are an absolute requirement for exsheathment. Exsheathment of four abomasal species was investigated in both the presence and absence of CO2, in either rumen fluid (cow or sheep) or buffer (standard or enriched). Exsheathment of Ostertagia ostertagi, Teladorsagia circumcincta and Ostertagia leptospicularis was observed in CO2-depleted rumen fluid and enriched buffer (respectively 46%, 22% and 15% in rumen fluid and 28% 18% and 26% in enriched buffer after 24 h). The level of this response was dependent on the species as well as the medium, and exsheathment was significantly higher in the presence of CO2. For Haemonchus contortus, exsheathment could only be achieved under CO2-saturated conditions. In conclusion, even though these parasite species exsheath in the same environment, there were significant differences in the minimal requirements to trigger their exsheathment. Some abomasal species were capable of exsheathment in the absence of CO2, which is likely facilitated by cofactors present in the rumen fluid and/or enriched buffer.


Assuntos
Dióxido de Carbono/metabolismo , Ostertagia/crescimento & desenvolvimento , Ostertagia/metabolismo , Trichostrongyloidea/crescimento & desenvolvimento , Trichostrongyloidea/metabolismo , Animais , Bovinos , Doenças dos Bovinos/parasitologia , Haemonchus/fisiologia , Larva , Rúmen/química , Rúmen/parasitologia , Ovinos , Doenças dos Ovinos/parasitologia , Temperatura
15.
J Anim Sci ; 96(11): 4868-4881, 2018 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-30085144

RESUMO

Active plant metabolites (APM) are recognized as modifiers of ruminal microbial fermentation including methanogenesis and biohydrogenation of fatty acids (FA). Coleus amboinicus Lour. leaves (CAL) are rich in several APM, which could serve as ruminal fermentation modulators. A phytochemical analysis showed that CAL contain phenolic acids (10.4 mg·g-1 dry matter [DM]; high in rosmarinic acid), flavonoids (2.6 mg·g-1 DM), diterpenes (2 mg·g-1 DM), and linolenic acid (35.4 g (100 g)-1 FA). This study aimed to investigate the effect of CAL on ruminal methanogenesis and biohydrogenation as well as basic fermentation characteristics and microbial populations. The in vitro experiment was carried out using Hohenheim gas test system with 40 mL of buffered ruminal fluid incubated for 24 h at 39 °C in anaerobic conditions. Approximately 400 mg (DM basis) of total mixed ration (TMR) was used as a control substrate and the CAL substrate was used at doses of 10, 20, 40, and 80 mg DM replacing equal amounts of TMR. Addition of CAL decreased methane production up to 30% linearly as the amount of CAL increased (P < 0.05). In vitro dry matter digestibility and ammonia tended to increase with increasing doses of CAL. Concentration of total volatile fatty acids was not affected by the CAL although there appeared to be a minor positive linear trend; however, acetate, butyrate, and isobutyrate proportion increased quadratically (P < 0.001). CAL tended to linearly increase α-linolenic acid and conjugated linoleic acid as well as increased stearic acid concentration in buffered ruminal fluid. CAL particularly increased total protozoa and bacterial populations during fermentation, but inhibited methanogens. It is concluded that the CAL may be promising to be used as a feed additive to decrease methanogenesis as well as biohydrogenation of FA in the rumen.


Assuntos
Coleus/química , Suplementos Nutricionais , Ácidos Graxos/metabolismo , Metano/metabolismo , Preparações de Plantas/farmacologia , Animais , Dieta/veterinária , Digestão/efeitos dos fármacos , Ácidos Graxos Voláteis/análise , Feminino , Fermentação/efeitos dos fármacos , Hidrogenação , Hidroxibenzoatos/análise , Folhas de Planta/química , Preparações de Plantas/química , Rúmen/metabolismo , Rúmen/microbiologia , Rúmen/parasitologia
16.
J Microbiol Methods ; 152: 186-193, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30142357

RESUMO

Rumen protozoa, primarily ciliates, are one of the important groups of strictly anaerobic microbes living in the rumen. Despite their ubiquitous occurrence in the rumen and significant contribution to host animals, it is still poorly understood why they live only in the rumen and similar environment. Because rumen protozoa require strict anaerobic conditions to sustain their viability and grow, only a few laboratories equipped with protozoology expertise and anaerobic facilities can grow rumen protozoa in laboratory. Also for the same reason, only a few species have been grown and maintained as laboratory cultures for research. Prompted by a recent study, we hypothesized that anaerobic rumen protozoa could also be cultivated aerobically if antioxidants were included in the media. Indeed, our experiments showed that the cultures of both Entodinium caudatum and Epidinium caudatum, two major rumen protozoal species, could be cultured successfully in aerobic media supplemented with ascorbic acid, glutathione and α-ketoglutarate as antioxidants. Anaerobic fermentation was maintained through the fermentation characteristics and microbial populations were altered to some extent under aerobic conditions. The antioxidants also enhanced the revival of cryopreserved stock cultures of both rumen protozoal species. The results of this study may facilitate and promote future research in which rumen protozoa need to be cultured in laboratory.


Assuntos
Técnicas de Cultura de Células/métodos , Cilióforos/crescimento & desenvolvimento , Rúmen/parasitologia , Anaerobiose , Animais , Antioxidantes , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Cilióforos/classificação , Meios de Cultura/química , Fermentação , Genes de Protozoários , RNA Ribossômico 16S/genética , Rúmen/microbiologia , Ruminantes/microbiologia , Ruminantes/parasitologia
17.
J Anim Physiol Anim Nutr (Berl) ; 102(5): 1111-1119, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29920791

RESUMO

Vitamin E (Vit. E) is discussed to influence ruminal biohydrogenation. The objective of this study was to investigate the influence of a Vit. E supplementation on rumen fermentation characteristics, ruminal microbial protein synthesis as well as ruminal organic matter fermentation. Furthermore, we aimed to investigate the influence of Vit. E supplementation on short-chain fatty acids (SCFA) and protozoa concentrations in the rumen and, in addition, on transfer rates of middle-chain and long-chain fatty acids into the duodenum in lactating dairy cows. Eight rumen and duodenum fistulated German Holstein cows were assigned to either a group receiving 2,327 IU/d Vit. E (138.6 IU/kg DM DL-α-tocopherylacetate; n = 4) or a control group (23.1 IU/kg DM; n = 4). Neither ruminal protein synthesis nor organic matter fermentation was influenced by treatment. Vit. E did not act on the concentrations of short-chain fatty acids and protozoa in rumen fluid. Duodenal flow of C13:0 (1.3 versus 0.2 g/d, p = 0.014) and iso-C14:0 (1.0 versus 0.5 g/d, p = 0.050) was higher in the Vit. E group. We observed a trend for higher duodenal flows for C12:0 (1.6 versus 0.9 g/d, p = 0.095) and anteiso-C15:0 (12.2 versus 8.9 g/d, p = 0.084). Transfer rate of C12:0 tended to be higher in the Vit. E group (125.61 versus 73.96, p = 0.082). No other transfer rates were affected by treatment. Further studies are necessary to investigate the influence of Vit. E on rumen microbiota and their fatty acid production as well as on the impact of different doses of Vit. E supplementation on variables of protein synthesis efficiency.


Assuntos
Ácidos Graxos/metabolismo , Fermentação/efeitos dos fármacos , Rúmen/metabolismo , Rúmen/parasitologia , Vitamina E/farmacologia , Animais , Bovinos , Dieta , Duodeno/metabolismo , Ácidos Graxos Voláteis/metabolismo , Feminino , Lactação , Leite
18.
Parasitol Int ; 67(5): 533-537, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29758277

RESUMO

More than 70 species of the Superfamily Paramphistomoidea, have been identified in ruminants in different parts of the world. Most are pathogenic, causing amphistomosis. Adult flukes within this family have a predilection for the forestomach (rumen) or bile duct of the liver, where they may cause epithelial damage. Identification of adult Paramphistomum, Calicophoron, Gastrothylax and Fischoederius at the species level based on morphology requires specialised expertise, whereas molecular genetic marker analysis is more precise and transferable. In the present study, we performed molecular characterisation of twenty seven adult flukes collected from the forestomachs of buffalo, cattle and goats in the Punjab province of Pakistan. PCR and sequencing of the ITS-2 rDNA region revealed a single haplotype in all cases. Phylogenetic comparison of P. epiclitum ITS2-rDNA sequences with those from other Paramphistomum, Calicophoron, Gastrothylax and Fischoederius species was performed to assess within and between species variation and validate the use of ITS-2 rDNA as a robust species-specific marker for P. epiclitum identification. This work provides a validated species-specific marker of P. epiclitum and the first report of this parasite species from Pakistan. The results of this study also have implications for the diagnosis and control of rumen flukes in the region and the need for accurate species identification to understand parasite distribution and population genetics.


Assuntos
Paramphistomatidae/genética , Rúmen/parasitologia , Infecções por Trematódeos/veterinária , Animais , Búfalos/parasitologia , Bovinos/parasitologia , Doenças dos Bovinos/parasitologia , DNA Intergênico/genética , Marcadores Genéticos , Doenças das Cabras/parasitologia , Cabras/parasitologia , Haplótipos , Paquistão , Paramphistomatidae/isolamento & purificação , Contagem de Ovos de Parasitas , Filogenia , Reação em Cadeia da Polimerase
19.
J Anim Physiol Anim Nutr (Berl) ; 102(3): 662-669, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29363190

RESUMO

This study was conducted to evaluate the effect of supplying two levels of Acacia nilotica (A. nilotica) pods to rations of sheep on nutrient digestibility, nitrogen balance and rumen liquor parameters (pH, total protozoa count, protein concentration and enzymes activity). Twelve mature rams (50 ± 1.25 kg B.W.) were distributed into three groups, each with four rams. Animals in group one were considered as a control which fed a basal diet, consisting of concentrate mixture and Egyptian clover. The second group and the third one received the same basal diet with supplying the concentrate mixture by 1.5% and 3.0% of A. nilotica pods meal respectively. The experiment lasted for 3 weeks. It was found that supplementation of A. nilotica pods to the concentrate mixture at a rate of 1.5% and 3.0% significantly improved the total feed intake compared to the control. The digestibility of dry matter and crude fibre was significantly reduced with A. nilotica supplements, whereas the digestibility of crude protein was significantly improved. All of nitrogen intake and N-retained were significantly increased in rams fed on concentrates with 1.5% and 3.0% A.  nilotica pods when compared to the control. The pH of ruminal fluid was not affected by the dietary treatments. Nevertheless, the total rumen protozoa count was significantly decreased in A. nilotica pods supplemented groups. Also, the rumen protein concentration and the ruminal enzymes activity, especially α-amylase, cellulase and protease, were lower in A. nilotica pods supplemented treatments. In conclusions, inclusion of low levels of A. nilotica pods (1.5% and 3.0%) in the concentrates can be used as a natural protein protectant in ruminants by forming tannin protein complexes in the rumen to maximize the amino acids available in the lower digestive tract. Also, these levels can increase the protein digestibility as well as the N-retained in the body .


Assuntos
Acacia , Ração Animal/análise , Dieta/veterinária , Rúmen/parasitologia , Ovinos/fisiologia , Animais , Digestão/fisiologia , Masculino , Nitrogênio/metabolismo , Rúmen/enzimologia
20.
J Dairy Sci ; 101(2): 1060-1073, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29174145

RESUMO

The objective of this study was to apply digital imaging to improve quantification of rumen protozoal biomass and distinguish treatment differences in cell motility and volume among ruminal protozoa. Observations of protozoa in rumen fluid treated with essential oils (CinnaGar, CIN; Provimi North America, Brookville, OH) or an ionophore (monensin, MON) indicated possible cell shrinkage. We hypothesized that MON would decrease protozoal motility and interact with CIN on cell volume. In addition, we hypothesized that analysis of still frames from video of swimming protozoa would improve volume prediction accuracy. Flocculated rumen fluid was incubated in batch culture dosed with N-free feed only (control), MON, CIN, or a combination of MON+CIN. Samples were taken at 0, 3, or 6 h post-treatment and wet-mounted on a microscope fitted with a high-definition camera. At 3 h post-inoculation, there was a treatment interaction for average speed such that CIN attenuated the effect of MON, with treatment means of 243, 138, 211, and 183 µm/s for control, MON, CIN, and MON+CIN, respectively. At 6 h post-inoculation, MON decreased average speed by 79.2 µm/s compared with the main effect mean without MON. We measured both minimum and maximum diameters (depth and width, respectively) perpendicular to the longitudinal axis of swimming protozoa, yielding a 3-dimensional estimate of protozoal volume. The ellipsoid formula (4/3)πabc, where a = 1/2 length, b = 1/2 width, and c = 1/2 depth, was compared with previously published volume estimations using genera-specific coefficients (genera-specific coefficient × length × width2). Residuals (genera-specific coefficients - ellipsoid) were plotted against predicted (ellipsoid) and centered to the mean (Xi-x¯) to evaluate both mean and slope biases. For Entodinium spp., Y = 0.248 (±0.037) (Xi - 7.98 × 104) + 1.97 × 104 (±1.48 × 103); n = 100; r2 [coefficient of determination (squared correlation coefficient)] = 0.31, with significant slope and mean biases. For family Isotrichidae, Y = -0.124 (±0.068) (Xi - 2.54 × 106) - 1.21 × 104 (±4.86 × 104); n = 32; r2 = 0.10, where slope tended to be different from zero but with no mean bias. For Epidinium spp., Y = 0.375 (±0.056) (Xi - 2.45 × 105) + 6.65 × 104 (±0.28 × 104); n = 64; r2 = 0.43, with both mean and slope biases. The present regression analyses demonstrate that the genera-specific coefficient-based method more likely overestimates volume for Entodinium and Epidinium than for the teardrop-shaped Isotrichidae. Based on simulations derived from previous literature reporting treatments that depress protozoal populations or among-animal changes in protozoal population structures, our proposed ellipsoid method offers potential to advance the prediction of treatment effects on protozoal volume and to shift focus from the number of cells present to the diversity, function, and biomass of protozoa under various treatment conditions.


Assuntos
Doenças dos Bovinos/parasitologia , Infecções por Cilióforos/veterinária , Cilióforos/fisiologia , Microscopia de Vídeo/métodos , Rúmen/parasitologia , Animais , Antiprotozoários/administração & dosagem , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Tamanho Celular/efeitos dos fármacos , Cilióforos/efeitos dos fármacos , Infecções por Cilióforos/tratamento farmacológico , Infecções por Cilióforos/parasitologia , Concentração de Íons de Hidrogênio , Monensin/administração & dosagem , América do Norte
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