Bovine Feces Research Articles

Fecal non-aureus Staphylococci are a potential cause of bovine intramammary infection

The presence of non-aureus staphylococci (NAS) in bovine rectal feces has recently been described. Similar to other mastitis causing pathogens, shedding of NAS in the environment could result in intramammary infection. The objective of this study was to investigate whether NAS strains present in feces can cause intramammary infection, likely via teat apex colonization. During a cross-sectional study in 5 dairy herds, samples were collected from the habitats quarter milk, teat apices, and rectal feces from 25%, 10%, and 25% of the lactating cows, respectively, with a cow serving as the source of one type of sample only. Samples from clinical mastitis cases were continuously collected during the 1-year study period as well. The 6 most prevalent NAS species, Staphylococcus (S.) chromogenes, S. cohnii, S. devriesei, S. equorum, S. haemolyticus, and S. hominis, were further subtyped by random amplification of polymorphic deoxyribonucleic acid polymerase chain reaction (RAPD-PCR), when the same NAS species was present in the same herd in the three habitats. For S. chromogenes, S. cohnii, S. devriesei, and S. haemolyticus, the same RAPD type was found in rectal feces, teat apices, and quarter milk, indicating that fecal NAS can infect the mammary gland. For S. hominis and S. equorum, we were unable to confirm the presence of the same RAPD types in the three habitats.

Recovery and seeds germination of four important tropical forage legumes in bovine dung

This experiment was conducted to evaluate the recovery and survival of tropical forage legume seeds (butterfly pea, stylosanthes, perennial soybean and macrotiloma) subjected through the digestive tract of cattle and to evaluate the germination of the seeds in feces of cattle. A 50 g of seed mixed with 150 g of mineral supplement offered to the cattle. Each cattle was considered as repetition. The bovine feces collected up to 60 hours after eating the seeds, where the seeds separated by sieves, using water, gloves procedure and tweezers. Later, they counted and divided into intact and swollen. For the germination test of the recovered seeds, 75 seeds per replicate were used (25 in natura, 25 intact and 25 swollen seeds, respectively). For the germination test in fecal plates, the bovine feces collected between 12 and 30 hours after eating the seeds. After 120 days, we evaluated the total amount of plants emerged within the study period (between 12 and 30 hours) and the average number of emerged plants per fecal signs. The results showed that cattle are facilitating the dispersion of butterfly pea, macrotyloma and perennial soybean, but not stylosanthes. The highest survival as the average number of germinated plants in fecal plates achieved by macrotyloma, followed by butterfly pea species and perennial soybean. The cattle were efficient in the pasture colonization process, due to the considerable dispersion of the seeds. This dispersion was not observed for seeds of stylosanthes.

First Record of Sarcophagidae (Diptera) Species in Seasonally Dry Tropical Forests (Caatinga), with Notes on Their Attractiveness to Decomposing Substrates1

Sarcophagidae or flesh flies are a highly diverse family of Diptera whose geographical distribution in the Neotropics is still poorly understood. We performed a two-year survey of flesh flies in fragments of Seasonally Dry Tropical Forests (SDTF) in Northeastern Brazil (Caatinga) using three types of substrate as baits: bovine spleen, fish and feces. Thirty-one species are registered, of which 13 are reported for the first time in SDTF. The highest diversity and abundance of flesh flies was associated with spleen. A strong female bias was observed in all samples. We discuss the medical, veterinary and forensic relevance and aspects of the synanthropy of the species registered for the first time in the Caatinga.

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The document associated with this DOI has been withdrawn.

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The document associated with this DOI has been withdrawn.

Potential sources of infection with selected zoonotic agents in the veterinary work environment - pilot studies.

The problem of occupational biohazards is very important, especially in the field of agriculture and in human and veterinary medicine. The aim of the study was to determine the potential sources of infection in veterinary professionals with selected zoonotic agents, including: Toxoplasma gondii, Giardia duodenalis, Leptospira spp., Cryptosporidium spp. and Coxiella burnetii. A total of 50 air samples from barns, piggeries and veterinary surgeries were examined for the presence of Leptospira spp. and C. burnetii DNA. Serum samples of 86 pigs and 80 cows were tested for the presence of antibodies to Leptospira spp. and to phase I and II C. burnetii antigens. Serum of 70 cats were tested for the presence of antibodies to T. gondii and 65 samples of cat faeces for the presence of T. gondii oocysts. The presence of G. duodenalis and Cryptosporidium spp. were examined in 50 of dog faeces and 50 of bovine faeces samples. DNA of Leptospira spp. was detected in 2 air samples from the piggeries (4%). C. burnetii DNA was not found in any sample. Anti-Leptospira spp. antibodies were detected in 51 (59.3%) of examined pigs. Neither anti-Leptospira spp. nor anti-C. burnetii antibodies were found among samples of bovine serum. Anti-T. gondii antibodies was found in 52 cat serum samples (74.3%). Among samples of cat faeces, no T. gondii oocysts were detected. In one sample of cattle stool (2%), G. duodenalis was detected and in another (2%) - Cryptosporidium spp. G. duodenalis was detected in 7 samples (14%) and Cryptosporidium spp. in 2 samples (2%) of dog faeces. The results of this study demonstrate the potential risk of infection with Leptospira spp. in veterinarians working with pigs. Veterinarians could be also be at risk of infection with T. gondii and G. duodenalis.

Dung decomposition of cattle grazing from mixed pastures of Signalgrass (Brachiaria decumbens Stapf.) and tree legumes

The mineralization rate of ruminant manure may influence the fertilization management of pastures. This study aimed to evaluate feces decomposition of heifers grazing signalgrass (Brachiaria decumbens Stapf.) fertilized or not with N, or intercropped with legumes in the dry forest region. Two experiments were conducted; the first one was a CRD that evaluated the evolution of CO2 from a mixture of soil and feces (10:1) during 22 days of incubation in a hermetically sealed bucket with a solution of NaOH 0.5 mol L-1. The second one was a RCBD that evaluated the in situ decomposition of feces in nylon bags in time periods 4, 8, 16, 32, 64, 128 and 256 days after incubation above ground. The single negative exponential mathematical model was adequate (P ≤ 0.0001) to quantify the CO2 evolution of the mixture of soil and feces, indicating that 78% of CO2 was released at the beginning of the incubation, especially for the feces collected in the signalgrass pastures intercropped with Gliricidia sepium (Jacq.) Kunth ex Walp. (gliricídia). After the first 5 days, CO2 evolution was more stable. Remaining biomass in the litterbag along decomposition fitted the single negative exponential model (P < 0.001). Greater relative decomposition rate (k) of bovine fecal biomass occurred for the N-fertilized signalgrass treatment (k = 0.0031 g g-1 day-1) and a lesser rate for the treatment intercropped with Mimosa caesalpiniifolia Benth. (sabiá) (k = 0.0018 g g-1 day-1). Nitrogen fertilization in signalgrass pasture favored the decomposition of bovine feces at the end of 256 days of incubation.

Application value of a dynamic automatic identification system in Schistosoma japonicum miracidium hatching test

To evaluate the value of a dynamic automatic identification system in routine miracidium hatching test with nylon gauzes. Different quantities of fresh Schistosoma japonicum eggs were added to bovine fecal samples and divided into the low-infection group, medium-infection group and high-infection group, while the bovine feces without S. japonicum eggs served as negative controls. The detection efficiency and accuracy were compared between the identification system and manual detection in different groups. The identification system can automatically identify S. japonicum miracidium. The detection rate and efficiency of S. japonicum miracidium in bovine fecal samples were both higher by using the identification system than by manual detection. Notably in the low-infection group, the identification system had a significantly higher rate of detection of S. japonicum miracidium than manual detection (χ2 = 10.769, P = 0.002). The identification system completed the detection of bovine fecal samples in the field within 1 min. The dynamic automatic identification system may effectively improve the detection efficiency and accuracy of routine miracidium hatching test with nylon gauzes, and it may replace manual detection to be used in the field schisotsomiasis examinations and related researches.

Mcr-1 encoding colistin resistance in CTX-M-1/CTX-M-15- producing Escherichia coli isolates of bovine and caprine origins in Tunisia. First report of CTX-M-15-ST394/D E. coli from goats

The objective of this study was to isolate and characterize ESBL-producing Escherichia coli (ESBL-EC) from raw bovine and caprine milk samples, as well as from bovine faeces in Tunisia. Therefore, 120 bovine faecal samples and 9 caprine raw milk samples were collected from 2 extensive dairy-cow-farms and 5 ovine farms, respectively. In addition, 94 raw bovine milk samples, from containers and holding tanks from 50 small public-markets in the North of Tunisia, were processed for the isolation of cefotaxime-resistant E. coli (CTXR). Antimicrobial susceptibility testing was carried out by disc-diffusion/broth-microdilution methods. The presence of genes encoding ESBL, as well as those encoding colistin (mcr-1 to 5 genes)- sulfonamide-, tetracycline-, gentamicin-, quinolone and chloramphenicol-resistance and class 1 integrons were tested by PCR (and sequencing in some cases). ESBL-EC isolates were further characterized by phylogrouping and MLST/PFGE typing. Eight samples (3.6%) contained ESBL-EC isolates (3/2 from raw bovine/goat milk and 3 from cattle faeces) and one isolate/sample was characterized. Four ESBL-EC isolates, all of bovine origin (3 faeces/1 milk), were resistant to colistin (MIC: 8–16 μg/ml), harboured the mcr-1 gene and carried IncP- and IncFIB-type plasmids. The 8 ESBL-EC strains had the following characteristics: a) bovine faeces: mcr-1/CTX-M-1/D-ST1642 (3 strains); b) raw milk: mcr-1/CTX-M-1/A-ST10 (1 strain); CTX-M-15/B1-ST394 (3 strains), and CTX-M-15/A-ST46 (1 strain). Most of bovine ESBL-EC isolates were multidrug-resistant (4/5). Our results showed that ESBL-EC were detected in bovine and caprine samples (CTX-M-1/CTX-M-15 producers), being some of them colistin-resistant (associated with mcr-1 gene), and they belonged to international clonal lineages.

Effectiveness of infectious source control after transmission control of schistosomiasis in Eryuan County of Yunnan Province

To assess the effectiveness of infectious source control at various stages of transmission control of schistosomiasis in Eryuan County of Yunnan Province, so as to provide insights into the further monitoring and management of infectious sources of schistosomiasis. Wenbi Village, a plateau subtype region and Qiandian Village, a mountain subtype region in Eryuan County were selected as the study areas. The species, schistosome infection and transmission capability of infectious sources were investigated in Wenbi and Qiandian villages in 2011 and 2018 and were compared. Schistosome infections were detected in human, bovine, horse, dog and mouse in Qiandian Village in 2011, with positive rates of 1.19%, 0.91%, 1.25%, 3.13% and 0.95%, respectively, and human, bovine and dog were found to have schistosome infections, with positive rates of 0.76%, 1.66% and 9.30%, respectively. However, no infections were identified in human, bovine, horse, pig, dog, sheep or mouse in these two villages in 2018. A total of 731 wild feces were collected in both villages in 2011. In Qiandian Village, horse, bovine and dog feces accounted for 34.40%, 29.80% and 20.20% of all fecal samples, and dog and horse feces were found to have schistosome infections (11.94% and 6.90% positive rates, respectively). In Wenbi Village, dog, bovine and human feces accounted for 44.59%, 39.83% and 14.29% of all fecal samples, and dog, human and bovine feces were found to have schistosome infections (16.95%, 9.52% and 2.33% positive rates, respectively). In 2018, a total of 204 wild feces were collected in both villages, and no schistosome infections were identified. Sheep, dog and bovine feces accounted for 36.27%, 33.33% and 27.45% of all fecal samples in Qiandian Village, and dog, bovine and human feces accounted for 72.55%, 11.76% and 10.78% of all fecal samples in Wenbi Village. A remarkable achievement has been obtained in the control of infectious sources of schistosomiasis in Eryuan County, and the role of human and bovine as the major infectious sources of schistosomiasis has been effectively controlled. In the future, the integrated strategy with emphasis on the control of infectious sources should be intensified, and the management of reservoir hosts including dog and mouse should be strengthened.

Occurrence of Escherichia coli O157:H7 in Pest Flies Captured in Leafy Greens Plots Grown Near a Beef Cattle Feedlot

Leafy greens are leading vehicles for Escherichia coli O157:H7 foodborne illness. Pest flies can harbor this pathogen and may disseminate it to produce. We determined the occurrence of E. coli O157:H7-positive flies in leafy greens planted up to 180 m from a cattle feedlot and assessed their relative risk to transmit this pathogen to leafy greens. The primary fly groups captured on sticky traps at the feedlot and leafy greens plots included house flies (Musca domestica L.), face flies (Musca autumnalis L.), stable flies (Stomoxys calcitrans L.), flesh flies (family Sarcophagidae), and blow flies (family Calliphoridae). E. coli O157:H7 carriage rates of house, face, flesh, and blow flies were similar (P > 0.05), ranging from 22.3 to 29.0 flies per 1,000 flies. In contrast, the carriage rate of stable flies was lower at 1.1 flies per 1,000 flies (P < 0.05). Differences in carriage rates are likely due to the uses of fresh bovine feces and manure by these different pest fly groups. E. coli O157:H7 carriage rates of total flies did not differ (P > 0.05) by distance (ranging from 0 to 180 m) from the feedlot. Most fly isolates were the same predominant pulsed-field gel electrophoresis types found in feedlot surface manure and leafy greens, suggesting a possible role for flies in transmitting E. coli O157:H7 to the leafy greens. However, further research is needed to clarify this role and to determine set-back distances between cattle production facilities and produce crops that will reduce the risk for pathogen contamination by challenging mechanisms like flies.

English

Escherichia coli present the ampC naturally, and the observation of phenotypical resistance to cefoxitin is related to this gene deregulation. Mutations in the regulatory region in ampC cause exaggerated expression. The most frequent alterations in the E. coli AmpC promoter/attenuator leading to this overexpression is described at the positions: -88, -82, -42, -18, -1 and +58. Mastitis studies were carried in Rio de Janeiro and Mato Grosso, Brazil. Two cefoxitin and amoxicillin-clavulanic acid-resistant E. coli from farms animals were unusually detected once these characteristics are not observed together in this species. The objective of this work was to determine if these isolates had a chromosomal gene mutation, determining AmpC hyperproduction. After DNA sequencing, mutations were observed at -88, -82, -73, -18, -1 and +58 positions, confirming the initially suspected AmpC hyperexpression. In Brazil, this is the first work to report E. coli hyperproducing this enzyme. &nbsp; Key words: Ampc attenuator, AmpC hyperproduction, ampC promoter, bovine feces, mastitic milk.

Dynamic Gene Network Analysis of Caco-2 Cell Response to Shiga Toxin-Producing Escherichia coli-Associated Hemolytic–Uremic Syndrome

Shiga toxin-producing Escherichia coli (STEC) O113:H21 strains are associated with human diarrhea and some strains may cause hemolytic–uremic syndrome (HUS). In Brazil, these strains are commonly found in cattle but, so far, were not isolated from HUS patients. Here, a system biology approach was used to investigate the differential transcriptomic and phenotypic responses of enterocyte-like Caco-2 cells to two STEC O113:H21 strains with similar virulence factor profiles (i.e., expressing stx2, ehxA, epeA, espA, iha, saa, sab, and subA): EH41 (Caco-2/EH41), isolated from a HUS patient in Australia, and Ec472/01 (Caco-2/Ec472), isolated from bovine feces in Brazil, during a 3 h period of bacteria–enterocyte interaction. Gene co-expression network analysis for Caco-2/EH41 revealed a quite abrupt pattern of topological variation along 3 h of enterocyte–bacteria interaction when compared with networks obtained for Caco-2/Ec472. Transcriptional module characterization revealed that EH41 induces inflammatory and apoptotic responses in Caco-2 cells just after the first hour of enterocyte–bacteria interaction, whereas the response to Ec472/01 is associated with cytoskeleton organization at the first hour, followed by the expression of immune response modulators. Scanning electron microscopy showed more intense microvilli destruction in Caco-2 cells exposed to EH41 when compared to those exposed to Ec472/01. Altogether, these results show that EH41 expresses virulence genes, inducing a distinctive host cell response, and is likely associated with severe pathogenicity.

Antimicrobial activity of essential oils extracted from clove and lemongrass against pathogenic bacteria isolated from bovine, swine and poultry feces

The objective was to verify the antibacterial activity of lemongrass and clove oils against Escherichia coli isolated from poultry feces, Staphylococcus aureus isolated from swine and poultry feces and Salmonella sp. isolated from swine and bovine feces. The antimicrobial activity was evaluated by the disc diffusion test using different concentrations of the oils: 160, 80, 40, 20 and 10 ?l ml -1. The multi-resistance of strains relative to that of conventional antimicrobials was also evaluated by the disk diffusion technique, using Multiple Antibiotic Resistance (MAR) index. The oils were characterized based on the results of chromatographic analyses, of which, analysis of lemongrass has been previously published (AZEVEDO et al., 2016). Lemongrass and clove essential oils presented citral and eugenol as the major compound, respectively. The concentrations of the essential oils had a significant effect on the extent of the growth inhibition halo and the coefficient of determination (r²) was higher than 0.80. Clove essential oil generated the largest zone of inhibition when tested against Escherichia coli and S. aureus from poultry feces and Salmonella sp. from the feces of swine, while lemongrass essential oil presented better results against S. aureus isolated from swine feces and Salmonella sp. from bovine feces. S. aureus and Salmonella sp. were multi-resistant to the antimicrobials tested. It is concluded that the essential oils tested have antimicrobial activity against bacteria isolated from bovine, swine, and poultry feces and that this activity is proportional to the concentration of oils and the microorganisms studied.

Depth-Dependent Response of Fecal Indicator Bacteria in Sediments to Changes in Water Column Nutrient Levels.

Concentrations of in bottom sediments can influence the assessment of microbial stream water quality. Runoff events bring nutrients to streams that can support the growth of in sediments. The objective of this work was to evaluate depth-dependent changes in populations after nutrients are introduced to the water column. Bovine feces were collected fresh and mixed into sediment. Studies were performed in a microcosm system with continuous flow of synthetic stream water over inoculated sediment. Dilutions of autoclaved bovine manure were added to water on Day 16 at two concentrations, and KBr tracer was introduced into the water column to evaluate ion diffusion. Concentrations of , total coliforms, and total aerobic heterotrophic bacteria, along with orthophosphate-P and ammonium N, were monitored in water and sediment for 32 d. Sediment samples were analyzed in 0- to 1-cm and 1- to 3-cm sectioned depths. Concentrations of and total coliforms in top sediments were approximately one order of magnitude greater than in bottom sediments throughout the experiment. Introduction of nutrients to the water column triggered an increase of nutrient levels in both top and bottom sediments and increased concentrations of bacteria in the water. However, the added nutrients had a limited effect on in sediment where bacterial inactivation continued. Vertical gradients of concentrations in sediments persisted during the inactivation periods both before and after nutrient addition to the water column.

Adverse effects of routine bovine health treatments containing triclabendazole and synthetic pyrethroids on the abundance of dipteran larvae in bovine faeces

Macrocyclic lactone treatments for livestock can have detrimental effects on the arthropod populations in livestock faeces. For the last twenty years, avoidance of these products has been a standard recommendation on livestock farms that are managed for wildlife by the Royal Society for Protection of Birds (RSPB). However, the continued decline in the populations of birds (in particular the red-billed chough Pyrrhocorax pyrrhocorax) that are dependent on dung invertebrates on islands in the Inner Hebrides of Scotland prompted us to investigate the effects of livestock treatments that are commonly used on these islands. We conducted a replicated field plot study over two years to quantify the effects of livestock treatments containing copper, deltamethrin and triclabendazole on invertebrate density in pooled, artificial faecal pats on the island of Islay. We found that the density of arthropod larvae was significantly reduced by the triclabendazole and deltamethrin treatments in both years and by as much as 86% when the treatments were combined. Copper-containing boluses did not consistently affect abundance of arthropod larvae. These results suggest that veterinary treatment of livestock might contribute to a reduction in the food supply of chough.

Comparative Metagenomics and Network Analyses Provide Novel Insights Into the Scope and Distribution of β-Lactamase Homologs in the Environment.

The β-lactams are the largest group of clinically applied antibiotics, and resistance to these is primarily associated with β-lactamases. There is increasing understanding that these enzymes are ubiquitous in natural environments and henceforth, elucidating the global diversity, distribution, and mobility of β-lactamase-encoding genes is crucial for holistically understanding resistance to these antibiotics. In this study, we screened 232 shotgun metagenomes from ten different environments against a custom-designed β-lactamase database, and subsequently analyzed β-lactamase homologs with a suite of bioinformatic platforms including cluster and network analyses. Three interrelated β-lactamase clusters encompassed all of the human and bovine feces metagenomes, while β-lactamases from soil, freshwater, glacier, marine, and wastewater grouped within a separate “environmental” cluster that displayed high levels of inter-network connectivity. Interestingly, almost no connectivity occurred between the “feces” and “environmental” clusters. We attributed this in part to the divergence in microbial community composition (dominance of Bacteroidetes and Firmicutes vs. Proteobacteria, respectively). The β-lactamase diversity in the “environmental” cluster was significantly higher than in human and bovine feces microbiomes. Several class A, B, C, and D β-lactamase homologs (blaCTX-M, blaKPC, blaGES) were ubiquitous in the “environmental” cluster, whereas bovine and human feces metagenomes were dominated by class A (primarily cfxA) β-lactamases. Collectively, this study highlights the ubiquitous presence and broad diversity of β-lactamase gene precursors in non-clinical environments. Furthermore, it suggests that horizontal transfer of β-lactamases to human-associated bacteria may be more plausible from animals than from terrestrial and aquatic microbes, seemingly due to phylogenetic similarities.

Next-generation sequencing analysis of bacterial flora in bovine protothecal mastitic milk and feces.

The aim of the present study was to evaluate the bacterial flora in the udder and intestinal environments in cows with and without protothecal mastitis. We used next-generation sequencing (NGS) analysis to identify 16S rRNA genes from bacterial flora present in 13 milk and 13 fecal samples from protothecal mastitic and healthy dairy cows in the Aichi region of Japan. Sequences associated with 5 species (Calothrix desertica, Corynebacterium simulans, Corynebacterium striatum, Empedobacter falsenii, and Rothia endophytica) showed the highest prevalence in samples of milk and feces from animals with protothecal mastitis. This range of species differed from those detected in the milk and feces from healthy cows.

Streptococcus agalactiae is not always an obligate intramammary pathogen: Molecular epidemiology of GBS from milk, feces and environment in Colombian dairy herds.

For many years Streptococcus agalactiae has been considered an obligate intramammary and strictly contagious pathogen in dairy cattle. However, recent reports of S. agalactiae isolation from extramammary sources have contradicted that premise. To gain further insight into the epidemiology of S. agalactiae infection in cattle, we examined its distribution and heterogeneity of strains in bovine milk, bovine feces, and the environment in Colombian dairy farms. First, a longitudinal study was conducted at herd level in 152 dairy herds. Bulk tank milk samples from each herd where collected twice a month for six months. A follow-up study with a cross sectional design at the cow level was conducted in a subset of 25 farms positive for S. agalactiae. Cow-level milk samples from 1712 lactatting cows and 1545 rectal samples were collected, as well as 120 environmental samples. Samples were used for S. agalactiae detection and genotyping using Multi Locus Sequence Typing. Results showed sporadic rather than repeated isolation of S. agalactiae from bulk tank milk in 40% of the positive herds, challenging the idea that S. agalactiae is a highly contagious pathogen causing chronic infections. S. agalactiae was isolated from rectal or environmental samples in 32% and 12% of cross-sectional study farms, respectively, demonstrating that the bacteria can survive in extramammary sources and that S. agalactiae is not an obligate intramammary pathogen. The same strain was isolated from rectal and bulk tank milk samples in eight farms, suggesting that fecal shedding is frequent, and contributes to the presence of S. agalactiae in bulk tank. High within-herd heterogeneity of strains was found, which is distinct from the situation in developed dairy industries. These new epidemiological findings should be considered to adjust surveillance and control recommendations for S. agalactiae.