Faecal Counts Research Articles

Space–time interactions between free-ranging red deer (Cervus elaphus) and grazing livestock on a fenced pasture

Livestock farming, which produces milk and meat, is practised by nearly all cultures. But in addition to this, grazing livestock fulfils other fundamentally important roles, one of which is the maintenance of open landscapes, which in turn provides habitat for many rare plant and animal species. Whether wild large herbivores, such as red deer (Cervus elaphus), benefit from the management of the open landscapes is unknown, and there are conflicting views on the nature of the interaction between livestock and wildlife. To investigate this question, we studied interactions in space and time between red deer and livestock in an 787 ha fenced pasture in eastern Germany. First, we used faecal pellet counts to determine whether red deer were using the fenced pasture and whether livestock were using the entire available pasture. Second, we fitted ten red deer, as well as eight Konik horses (Equus ferus caballus) and eight Heck cattle (Bos taurus) with GPS-collars. These data allowed us to perform analyses of space–time interactions. The faecal pellet data showed that red deer did indeed use the fenced pasture and that livestock used the entire extent of the pasture. Further, we verified a neutral space–time interaction between red deer and livestock within the pasture. Thus, there was no indication of a negative effect of the presence of livestock on red deer inside the pasture. For these reasons, we recommend that low-intensity grazing by cattle and horses can maintain open landscapes without detrimental effects on the use of the area by red deer.

Fecal bacteria in coastal lakes: An anthropogenic contamination or natural element of microbial diversity?

The effects of multiple stressors (salinity, temperature, nutrient and light availability or human activities) on the abundance of FIB (Escherichia coli and Enterobacteriaceae) were studied in nine lakes on the southern Baltic coast. Monitoring also included physicochemical factors and land use in the catchment. The results showed that FIB is a natural component of microbial community in coastal lakes, with its abundance varying temporally and spatially. The key factors predicting the abundance of E. coli and Enterobacteriaceae are the availability of ammonia nitrogen and water temperature. Regardless of the degree of connection with the sea, the sanitary condition of coastal lakes deteriorates during the summer months. Although agricultural use and residential development do not significantly affect fecal bacteria counts (PLS-R model), human pressure on the sanitary condition of lakes increases seasonally and is limited to the vicinity of tourist sites. The degree of contact with seawater regulates the size of the microbial community, but is not a factor that directly affects the presence of E. coli in the microbial community in coastal lakes. Monitoring the sanitary conditions of these ecosystems using sensitive, rapid survey methods based on hydrochemical and microbiological inventories, supported by GIS, is important for public health.

Impact of Selenium Nanoparticle-Enriched Lactobacilli Feeding Against Escherichia coli O157:H7 Infection of BALB/c Mice.

The effectiveness of selenium nanoparticle (SeNP)-enriched Lactiplantibacillus plantarum and Lactobacillus acidophilus was studied against Shiga toxin-producing Escherichia coli O157:H7 infection on the intestinal fragments and kidney tissue of BALB/c mice. Gut microbiota-targeted bacteria and E. coli O157:H7 counts were obtained by qPCR and PCR. Histology of ileum, colon, and kidney tissues and Stx secretions were analyzed until one-week post-infection. Mice fed with SeNP Lpb. plantarum in the preinfection feeding groups have lower E. coli O157:H7 counts and lower intestinal damage than those in the infected group. The lowest mean fecal probiotic counts were in the L. acidophilus group (7.61 log 10). In pretreatment groups of SeNP L. acidophilus and L. acidophilus, the mean counts of bacteria decreased to 104CFU/g by day 7. The lowest Stx copy number was demonstrated in SeNP Lpb. plantarum feeding groups' day 7 (P < 0.05). Feeding groups with SeNP Lpb. plantarum had significantly higher members of Lactobacilli in their fecal microbiota than the control group on day 7. It was clarified that Se-enriched Lpb. plantarum and L. acidophilus can be useful as a method of preventing STEC infections. The viability of STEC infection exposure to selenium-enriched Lactobacillus spp. was decreased more than for non-Se-enriched Lactobacillus spp.

A Potential Adhesin/Invasin STM0306 Participates in Host Cell Inflammation Induced by Salmonella enterica Serovar Typhimurium.

Salmonella enterica serovar typhimurium (S. Typhimurium) is a common Gram-negative foodborne pathogenic bacterium that causes gastrointestinal disease in humans and animals. It is well known that adhesins and invasins play crucial roles in the infection mechanism of S. Typhimurium. S. Typhimurium STM0306 has been denoted as a putative protein and its functions have rarely been reported. In this study, we constructed the STM0306 gene mutant strain of S. Typhimurium and purified the recombinant STM0306 from Escherichia coli. Deletion of the STM0306 gene resulted in reduced adhesion and invasion of S. Typhimurium to IPEC-J2, Caco-2, and RAW264.7 cells. In addition, STM0306 could bind to intestinal epithelial cells and induced F-actin modulation in IPEC-J2 cells. Furthermore, we found that STM0306 activated the nuclear factor kappa B (NF-κB) signaling pathway and increased the mRNA expression of pro-inflammatory cytokines such as IL-1β, TNF-α, as well as chemokine CXCL2, thus resulting in cellular inflammation in host cells. In vivo, the deletion of the STM0306 gene led to reduced pathogenicity of S. Typhimurium, as evidenced by lower fecal bacterial counts and reduced body weight loss in S. Typhimurium infected mice. In conclusion, the STM0306 of S. Typhimurium is an important adhesin/invasin involved in the pathogenic process and cellular inflammation of the host.

Evaluation of tributyrin supplementation in milk replacer on diarrhoea occurrence in preweaning Holstein calves

Neonatal calf diarrhoea is one of the most important health challenges in cattle herds causing substantial economic losses and antimicrobial use. Due to the raising problem of antimicrobial resistance, effective alternatives are urgently required, in line with European policies. The aim of this study was to evaluate the effect of tributyrin supplementation in milk replacer on diarrhoea, performance and metabolic status in preweaning Holstein calves. Twelve newborn calves, after colostrum administration, were randomly allotted in two experimental groups for 42 days: control (CTRL) fed milk replacer, tributyrin (TRIB) fed milk replacer supplemented with 0.3% of liquid tributyrin on milk powder weight. Calves BW was recorded on a weekly basis from day 7 to day 42, and feed intake was recorded daily to calculate zootechnical performance. Faecal consistency was assessed daily through the faecal score (0–3 scale; considering diarrhoea moderate = 2 and severe = 3). Faecal samples were collected weekly from rectal ampulla for microbiological analysis by plate counting method evaluating the number of total bacteria, lactic acid bacteria and coliform bacteria. On day 0 and day 42, individual blood samples were collected from jugular vein for metabolic profile analysis. Serum samples of day 42 were also evaluated for the antioxidant barrier using a colorimetric test, while glucagon-like peptide 2 and diamine oxidase concentrations were measured through immunoenzymatic assays. Tributyrin supplementation did not influence the zootechnical performance of calves over 42 days of trial. Diarrhoea frequency was significantly lower in TRIB compared to CTRL group (27.91 and 38.37%; P < 0.01) considering the whole experimental period. In particular, the major effect was observed for moderate diarrhoea in TRIB group that showed a significantly reduced frequency compared to CTRL (P < 0.01) thus suggesting a preventive effect of tributyrin. Faecal total bacterial, lactic acid and coliform bacteria counts did not show differences between groups. Urea serum concentrations tended to be lower in TRIB compared to CTRL, indicating an efficient utilisation of dietary protein. Antioxidant barrier and glucagon-like peptide 2 were comparable between CTRL and TRIB on day 42. Diamine oxidase concentrations were significantly decreased in TRIB compared to CTRL group after 42 days of trial (P < 0.01), suggesting a higher gut epithelial integrity probably due to lower diarrhoea frequency and the nourish effect of tributyrin on enterocytes. In conclusion, tributyrin could be considered as a valuable bioactive feed additive to decrease the neonatal diarrhoea occurrence and support intestinal integrity in preweaning calves.

Live and pasteurized Akkermansia muciniphila decrease susceptibility to Salmonella Typhimurium infection in mice

IntroductionThe gut microbiome is vital for providing resistance against colonized pathogenicbacteria. Recently, specific commensal species have become recognized as important mediators of host defense against microbial infection by a variety of mechanisms. ObjectivesTo examine the contribution of live and pasteurized A. muciniphila to defend against the intestinal pathogen Salmonella Typhimurium in a streptomycin-treated mouse model of infection. MethodsC57B6J mice were pretreated with phosphate-buffered saline (PBS), live Akkermansia muciniphila (AKK), and pasteurized A. muciniphila (pAKK) for two weeks, then mice were infected by S. Typhimurium SL 1344. 16S rRNA-based gut microbiota analysis was performed before and after infection. Bacterial counts in feces and tissues, histopathological analysis, gut barrier-related gene expression, and antimicrobial peptides were examined. Co-housing was performed to examine the role of microbiota in the change of susceptibility of mice to infection. ResultsAKK and pAKK markedly decreased Salmonella fecal and systemic burdens and reduced inflammation during infection. Notably, further characterization of AKK and pAKK protective mechanisms revealed different candidate protective pathways. AKK promoted gutbarrier gene expression and the secretion of antimicrobial peptides, and co-housing studies suggested that AKK-associated microbial community played a role in attenuating infection. Moreover, pAKK had a positive effect on NLRP3 in infected mice. We verified that pretreatment of pAKK could promote the expression of NLRP3, and enhance the antimicrobial activity of macrophage, likely through increasing the production of reactive oxygen (ROS), nitric oxide (NO), and inflammatory cytokines. ConclusionOur study demonstrates that live or pasteurized A. muciniphila can be effective preventive measures for alleviating S. Typhimurium-induced disease, highlighting the potential of developing Akkermansia-based probiotics or postbiotics for the prevention of Salmonellosis.

Pink1-/- Rats Demonstrate Swallowing and Gastrointestinal Dysfunction in a Model of Prodromal Parkinson Disease.

Early motor and non-motor signs of Parkinson disease (PD) include dysphagia, gastrointestinal dysmotility, and constipation. However, because these often manifest prior to formal diagnosis, the study of PD-related swallow and GI dysfunction in early stages is difficult. To overcome this limitation, we used the Pink1-/- rat, a well-established early-onset genetic rat model of PD to assay swallowing and GI motility deficits. Thirty male rats were tested at 4months (Pink1-/- = 15, wildtype (WT) control = 15) and 6months (Pink1-/- = 7, WT = 6) of age; analogous to early-stage PD in humans. Videofluoroscopy of rats ingesting a peanut-butter-barium mixture was used to measure mastication rate and oropharyngeal and pharyngoesophageal bolus speeds. Abnormal swallowing behaviors were also quantified. A second experiment tracked barium contents through the stomach, small intestine, caecum, and colon at hours 0-6 post-barium gavage. Number and weight of fecal emissions over 24h were also collected. Compared to WTs, Pink1-/- rats showed slower mastication rates, slower pharyngoesophageal bolus speeds, and more abnormal swallowing behaviors. Pink1-/- rats demonstrated significantly delayed motility through the caecum and colon. Pink1-/- rats also had significantly lower fecal pellet count and higher fecal pellet weight after 24h at 6months of age. Results demonstrate that swallowing dysfunction occurs early in Pink1-/- rats. Delayed transit to the colon and constipation-like signs are also evident in this model. The presence of these early swallowing and GI deficits in Pink1-/- rats are analogous to those observed in human PD.

Factors associated with stunting: gut inflammation and child and maternal-related contributors among under-five children in Hawassa City, Sidama Region, Ethiopia

BackgroundUnder-nutrition remains a major global public health challenge, particularly among children under the age of five. Among the manifestations of under-nutrition, stunting accounts for the larger proportion, which is associated with multiple factors. In Ethiopia, however, the link between intestinal inflammation and childhood stunting was not well investigated. Therefore, the present study aimed to determine the association between gut inflammation and childhood stunting.MethodA community-based cross-sectional study was conducted and a total of 82 children were included in the study. Anthropometric data were collected by measuring weight in underwear and without shoes with an electronic scale to the nearest 0.1 kg and their height in the Frankfort plane with a telescopic height instrument. Environmental risk factors for enteric bacterial exposure, access to improved sources of drinking water, and the presence of facilities for hygiene and sanitation conditions were assessed using a questionnaire. Gut inflammation was tested through fecal leukocyte count and each sample was stained with methylene blue. Stool samples were inoculated on MacConkey agar, Salmonella-Shigella agar, and Xylose Lysine Deoxycholate agar after enrichment with Selenite cystine broth and incubated at 37 °C for 18–24 h. Binary and multiple logistic regressions and Chi-square models were used to analyze the data.ResultData from the current study revealed that gut inflammation was (AOR: 5.28, 95% CI: 1.32–22.25) associated with stunting. On the other hand, children with reported diarrhea within the last week were 6 times more likely for the probability of being stunted (AOR: 6.21, 95% CI: 2.68–26.83). The findings of this study also demonstrated that children from a household with a family size of more than 5 members were three times more likely to be stunted than their counterparts (AOR: 3.21, 95% CI: 1.20 -10.13). Facts of the current study demonstrated that breastfeeding for 24 months and below was negatively associated (AOR: 0.3; 95% CI: -0.46-0.89) with gut inflammation. Detection of E.coli and Shigella species in the stool samples of children and Menaheria residents were positively associated with gut inflammation (AOR: 5.4, 95% CI: 1.32–22.25; AOR: 5, 95% CI: 1.47–24.21), respectively.ConclusionTherefore, there was a strong correlation between stunting and gastrointestinal inflammation. Moreover, stunting was associated with diarrhea, breastfeeding duration, residence, and family size. Similarly, intestinal inflammation was linked to residence, breastfeeding duration, and the prevalence of bacterial infections such as E. coli and Shigella species.

Epidemiological survey of gastrointestinal infections by gastrointestinal nematodes and coccidia in cattle in the semiarid region of Northeastern Brazil

The aim of the present study was to carry out an epidemiological survey on gastrointestinal infections by nematodes and coccidia in cattle herds in the semiarid region of Paraíba, northeastern Brazil. A total of 832 cattle were sampled, of both sexes and different ages, on 21 farms in 21 municipalities in the state of Paraíba, Brazil. Fecal samples were collected individually to perform eggs per gram (EPG) and oocysts per gram (OoPG) counts in feces, and to perform fecal cultures. To assess factors associated with nematode and coccidia infections, epidemiological survey forms were applied to the farmers. The total frequency of infections by nematodes was 59.6% (496/832) and it was 36.9% (307/832) for coccidia. The levels of infection by nematodes and coccidia were most frequently low (EPG &lt; 300; OoPG &lt; 1,000), in 72.4% (359/496) and 75.2% (231/307), respectively. The most frequent nematode genera was Haemonchus spp. (78.8%). The risk factors for nematode infections in cattle were beef production trait (odds ratio = 2.99) and extensive rearing system (odds ratio = 3.8); the factors for coccidia were age ≤ 12 months (odds ratio = 2.9) and body score between 1 and 2 (odds ratio = 5.2). These results prove that there are high frequencies of gastrointestinal infections by nematodes and coccidia in cattle herds in the semiarid region of Paraíba, with predominantly low levels of infection. However, given the importance of subclinical infections, adjustments to sanitary management measures can contribute to reduction of parasite indices and better performance among the animals, thus generating greater profitability for farmers.

The therapeutic effectiveness of thyme extract in naturally infected puppies with ascariasis

Aim: This study aimed to investigate the therapeutic efficacy of thyme extract in puppies naturally infected with ascarids.&#x0D; Methodology: The study consisted of 20 puppies of different sexes (males and females), 2-4 months old, naturally infected with ascarid. The experimental units were given an oral 20% concentration of thyme extract for 3 days, and faecal oocyst counts were conducted on the 0th, 1st, 2nd, 3rd, and 7th days starting from the day the treatment was started (day 0). Also, urea, creatinine, AST, ALT values in the blood were monitored on the 0th and 3rd days, together with daily clinical examination, to monitor possible toxic effects.&#x0D; Results: While the number of faecal egg counts in 2 puppies (10%) varied considerably, egg shedding in 8 (40%) of the treated puppies was zero. It was observed that in 10 of the puppies (60%), egg shedding was not completely zero. However, the oocyst counts decreased by 25% to 98.3%. It was also observed that the values of the measured blood biochemical values were within reference range and the puppies did no not show any clinical sign of toxicity during treatment.&#x0D; Conclusion and Recommendation: It was concluded that the thyme extract did not have any toxic effect in the puppies at the concentration studied, and it could be effective in the treatment of ascariasis.

Novel frontier in wildlife monitoring: Identification of small rodent species from fecal pellets using near-infrared reflectance spectroscopy (NIRS).

Small rodents are prevalent and functionally important across the world's biomes, making their monitoring salient for ecosystem management, conservation, forestry, and agriculture. There is a growing need for cost-effective and noninvasive methods for large-scale, intensive sampling. Fecal pellet counts readily provide relative abundance indices, and given suitable analytical methods, feces could also allow for the determination of multiple ecological and physiological variables, including community composition. In this context, we developed calibration models for rodent taxonomic determination using fecal near-infrared reflectance spectroscopy (fNIRS). Our results demonstrate fNIRS as an accurate and robust method for predicting genus and species identity of five coexisting subarctic microtine rodent species. We show that sample exposure to weathering increases the method's accuracy, indicating its suitability for samples collected from the field. Diet was not a major determinant of species prediction accuracy in our samples, as diet exhibited large variation and overlap between species. fNIRS could also be applied across regions, as calibration models including samples from two regions provided a good prediction accuracy for both regions. We show fNIRS as a fast and cost-efficient high-throughput method for rodent taxonomic determination, with the potential for cross-regional calibrations and the use on field-collected samples. Importantly, appeal lies in the versatility of fNIRS. In addition to rodent population censuses, fNIRS can provide information on demography, fecal nutrients, stress hormones, and even disease. Given the development of such calibration models, fNIRS analytics could complement novel genetic methods and greatly support ecosystem- and interaction-based approaches to monitoring.

Influence of Wormwood Seeds on Enzymatic and Non–enzymatic Activity in Blood of Broilers with Coccidiosis

Wormwood plants have been of great botanical and pharmaceutical interest and are employed in folk medicine for a long time. Wormwood has been used successfully for many years as a remedy against malaria and has been reported to exhibit antiprotozoal potential. Having in mind that, this study aimed to investigate the effects of wormwood seeds on enzymatic activity and non–enzymatic activity in broilers infected with coccidiosis. A biological experiment with broiler chickens was performed on a total of 360 Ross 308 broiler chickens. Chickens were divided into three experimental treatments with six replication each, respectively. Treatment I (WW1) was uninfected with coccidia oocyst and untreated, treatment II (WW2) was infected with coccidia oocyst and was kept untreated, while treatment III (WW3) was infected with coccidia oocyst and received wormwood seeds in feed in the quantity of 1.5%. Blood was collected for the evaluation of antioxidant status. Results of bloody diarrhea intensity of chickens have ranged from 0% (WW1; day 26th) to 99% (WW2; day 28th), respectively. Regarding the influence of dietary wormwood seeds on fecal oocyst counts obtained results showed the highest oocyst in treatment WW2 on the 30th day of the experiment (45115.7 oocyst number/g), which was significantly higher (p&lt;0.05) when compared to treatments WW3 (623.3 oocyst number/g), and WW1 (0 oocyst number/g) on the same experimental day. Our results indicate a significant (p&lt;0.05) increase in non-enzymatic activity and higher catalytic enzymatic activity in blood hemolysates of infected broilers. Based on our research it can be concluded that wormwood seeds can be used in feed for broilers as a prophylactic feed additive to prevent coccidia oocyst development.

English

The current study assessed the anticoccidial efficacy of chitosan nanoencapsulated bromelain (CNB) against coccidiosis in goats. Bromelain was extracted from the pineapple peels using standard methods while oral solution of CNB was prepared using standard manufacturing methods. The in vivo study was done on fifteen healthy male goats naturally infected with coccidia. The goats were divided into three groups consisting of three treatment groups (90 and 270 mg/Kg CNB, Diclazuril, 5 mg/Kg). The drugs were administered orally for 3 days. Fecal Oocyst Counts (FOC) determined using the modified McMaster technique. The goats were observed for clinical signs on daily basis while body weight was recorded weekly. The level of packed cell volume (PCV), aspartate aminotransferases (AST), alanine aminotransferases (ALT), urea, and creatinine were assessed weekly. At the end of the study, goats were euthanized and gross pathology and histopathology conducted. The results showed that at day 28 post-treatment there was a significantly reduction of FOC of 98.42 and 82.30% for Diclazuril and 270 mg/Kg treatment groups, respectively. The reduction of FOC percentage was significantly higher (p &le; 0.01) in Diclazuril group than that in 270 mg/Kg. During the monitoring period, there was no mortality or clinical signs observed in the goats. The PCV, AST, ALT, creatinine and urea were in normal ranges for goats. There were no pathological lesions on the goat organs. In conclusion, CNB had high anticoccidial efficacy and was safe for use in goats. Strategies to improve the efficacy of this potential drug should be further investigated. &nbsp; Key words: Anticoccidial efficacy, bromelain, chitosan, goat, nanoencapsulation.

A test of motion‐sensitive cameras to index ungulate densities: group size matters

AbstractThe use of species detection rates gathered from motion‐sensitive cameras as relative abundance indices (RAIs) could be a cost‐effective tool to monitor wildlife populations; however, validations based on comparisons with reference methods are necessary. We considered 3 ungulates, wild boar (Sus scrofa), roe deer (Capreolus capreolus), and fallow deer (Dama dama), and compared 2 different RAIs with independent indices of density obtained through feces counts across 3 summers (2019–2021) in a protected area of central Italy. We estimated the number of detections per day (RAIevents), and the number of individuals per day (RAIindividuals) from remote camera videos. Both indices were correlated with density estimates, yet only RAIindividuals correctly ranked interspecific densities. Values of RAIevents for the most abundant and gregarious ungulate (i.e., wild boar) were biased low and were lower than those of fallow deer. The uncertainty of RAIs was acceptable for the 2 most abundant study species (CVs ≤ 25%) but was greater for roe deer. At the intra‐specific level, density estimates and RAIs showed comparable but slight inter‐annual variation. Our results support the use of RAIs derived from motion‐sensitive cameras as a promising and cost‐effective tool to monitor ungulate populations, and researchers should incorporate group size into monitoring. We advocate the necessity of field tests based on comparison with locally reliable reference methods to validate the use of motion‐sensitive cameras.

Staphylococcus aureus sequence type (ST) 45, ST30, and ST15 in the gut microbiota of healthy infants — persistence and population counts in relation to ST and virulence gene carriage

Staphylococcus aureus colonizes the anterior nares, and also the gut, particularly in infants. S. aureus is divided into lineages, termed clonal complexes (CCs), which comprise closely related sequence types (STs). While CC30 and CC45 predominate among nasal commensals, their prevalence among gut-colonizing S. aureus is unknown. Here, 67 gut commensal S. aureus strains from 49 healthy Swedish infants (aged 3 days to 12 months) were subjected to multi-locus sequence typing. The STs of these strains were related to their virulence gene profiles, time of persistence in the microbiota, and fecal population counts. Three STs predominated: ST45 (22% of the strains); ST15 (21%); and ST30 (18%). In a logistic regression, ST45 strains showed higher fecal population counts than the others, independent of virulence gene carriage. The lower fecal counts of ST15 were linked to the carriage of fib genes (encoding fibrinogen-binding proteins), while those of ST30 were linked to fib and sea (enterotoxin A) carriage. While only 11% of the ST15 and ST30 strains were acquired after 2 months of age, this was true of 53% of the ST45 strains (p = 0.008), indicating that the former may be less fit for establishment in a more mature microbiota. None of the ST45 strains was transient (persisting < 3 weeks), and persistent ST45 strains colonized for significantly longer periods than persistent strains of other STs (mean, 34 vs 22 weeks, p = 0.04). Our results suggest that ST45 strains are well-adapted for commensal gut colonization in infants, reflecting yet-unidentified traits of these strains.

Portable near-infrared spectroscopy: A rapid and accurate blood test for diagnosis of Haemonchus contortus infection and for targeted selective treatment of sheep

Haemonchus contortus is the most prevalent and important gastrointestinal nematode (GIN) in small ruminants. Since it reduces the packed cell volume (PCV), causing anemia, early diagnosis can be used for targeted selective treatment (TST) of sheep, reducing antiparasitic drug use and anthelmintic resistance. This study aimed to predict PCV values through near-infrared spectroscopy (NIRS) and to develop a classification and diagnosis model of H. contortus infection using PCV values, eggs per gram of feces (EPG) counts and mean daily weight gain (DWG). A total of 1728 spectra were collected from blood samples of 216 lambs with a portable NIR spectroscope. In parallel, other parameters indicative of infection were measured: PCV by hematocrit, FAffa MAlan CHArt (FAMACHA) scores, EPG and DWG. To evaluate the relationship between NIRS spectra and the evaluated parameters, principal component analysis (PCA) was used for an exploratory analysis, regression by the partial least squares method (PLS) for the prediction of PCV values via NIRS, and PCA linear discriminant analysis (PCA-LDA) as a classification model for diagnosis. The absorption peaks in the NIRS region associated with the excitation of overtones of nitrogen-hydrogen (N-H) functional groups of proteins had a strong impact on the principal components (PCs), indicating that blood proteins, especially hemoglobin, can be estimated by the NIRS technique. The model for predicting PCV by PLS presented a standard error of prediction of 2.53, root-mean-square error of 2.48, and coefficient of determination of 0.84, indicating good correlation between the PCV values predicted by the model and the PCV obtained by hematocrit. The PCA-LDA model presented 93.33% sensitivity and 82.18% accuracy, both higher than those of the FAMACHA method, as was expected for resilient Morada Nova lambs. The multivariate models associated with the NIRS technique reported here can be used in the future as a quick and versatile tool for H. contortus infection diagnosis and TST application in lambs.

Probiotic for pathogen-specific Staphylococcus aureus decolonisation in Thailand: a phase 2, double-blind, randomised, placebo-controlled trial

Decolonisation is considered a valuable means to reduce Staphylococcus aureus infection rates. However, previous topical strategies targeting the nose or skin had little success, and oral antibiotic-based decolonisation is ill advised because of eradication of the microbiota and development of antibiotic resistance. We previously showed that the probiotic Bacillus subtilis significantly diminished S aureus at the main intestinal colonisation site via specific bacterial interaction in mice; in this study, we tested this probiotic approach to control S aureus colonisation in humans. We did a single-centre, phase 2, double-blind, randomised, placebo-controlled trial in adults from the Songkhla region of Thailand who were colonised by S aureus. Eligible participants were adults (aged ≥18 years) without history of intestinal disease, antibiotic treatment, or hospital admission within the previous 90 days. Participants were excluded if they were pregnant, breastfeeding, taking probiotics, or had diarrhoea. Participants were allocated (1:1) to groups by computer randomisation in blocks of four, and research coordinators were masked to group allocation. Participants received 250 mg of probiotic B subtilis MB40 or placebo once per day for 30 days and S aureus colonisation was determined after the last dose was received. The primary outcome was colonisation by S aureus (continuous, mean decrease in colony-forming-unit count) in the intestine (by faecal counts) and nares (by nasal swabs) after intervention (30-day regimen of B subtilis probiotic). This trial is registered with the Thai Clinical Trials Registry, TCTR20210128003. The trial was done between Jan 29 and June 30, 2021, with enrolment taking place from Jan 29 to April 6, 2021. 115 participants were colonised by S aureus, either in the intestine (n=84), nose (n=50), or both (n=19), and were randomly assigned to treatment (n=55) and placebo groups (n=60). Oral probiotic B subtilis resulted in significant reduction of S aureus in stool (96·8%; p<0·0001) and nose (65·4%; p=0·0002). There were no differences in adverse effects or significant microbiome changes between the intervention and placebo groups. B subtilis probiotic eliminated more than 95% of the total S aureus colonising the human body without altering the microbiota. This probiotic strategy offers several key advantages over presently used decolonisation strategies for potential use in people with chronic or long-term risk of S aureus infection. Furthermore, by establishing a defining role of the intestinal colonisation site, our findings call for revisiting fundamental notions about S aureus colonisation. National Research Council of Thailand and US National Institutes of Health.

Probiotic, Paraprobiotic, and Hydrolyzed Yeast Mixture Supplementation Has Comparable Effects to Zinc Oxide in Improving Growth Performance and Ameliorating Post-weaning Diarrhea in Weaned Piglets.

A total of 150 21-day-old weaned piglets [(Yorkshire × Landrace) × Duroc] were randomly assigned to 3 groups (CON, TRT1, TRT2) to evaluate the effects of dietary supplementation of probiotic, paraprobiotic, and hydrolyzed yeast mixture (PPY) on growth performance, nutrient digestibility, fecal bacterial counts, fecal calprotectin contents, and diarrhea rate in a 42-day experiment (phase 1: days 1-14; phase 2: days 15-42). There were 10 replicate pens per treatment with 5 pigs per pen (three gilts and two barrows). Pigs in CON were only provided with a basal diet. Pigs in TRT1 were provided with a basal diet + 3000mg/kg zinc oxide during phase 1 and a basal diet during phase 2. Pigs in TRT2 were provided with a basal diet + 200mg/kg probiotic (Saccharomyces cerevisiae boulardii) + 800mg/kg paraprobiotic (inactivated yeast strains of Saccharomyces cerevisiae and Cyberlindnera jadinii) + 10g/kg hydrolyzed yeast mixture during phase 1, and a basal diet + 100mg/kg probiotic + 400mg/kg paraprobiotic mixture during phase 2. Pigs in TRT1 and TRT2 were significantly heavier at day 14 and 42 than CON pigs. Growth rate during days 1-14, 15-42, and 1-42 and feed efficiency during days 1-14 were similarly affected by treatment while feed efficiency was significantly higher for TRT2 pigs between 15-42 and 1-42days. Moreover, nitrogen and energy digestibility in both TRT1 and TRT2 were higher than that in CON. During experimental periods, diarrhea rate in TRT1 and TRT2 was lower than that in CON. Therefore, we demonstrated that PPY supplementation had comparable effects as ZnO in improving growth performance and nutrient digestibility as well asameliorating post-weaning diarrhea in weaned piglets.

Getting a handle on rat familiarization: The impact of handling protocols on classic tests of stress in Rattus norvegicus.

Experimenter familiarization with laboratory rodents through handling prior to experimentation is an important practice in neurobehavioral research and is implicated in stress, study variability, and replicability. Unfortunately, different handling protocols have not been thoroughly examined. Determining optimal experimenter familiarization protocols is expected to reduce animal stress and thus improve welfare and data consistency. The impact of different handling protocols was determined through behavioral assessments (i.e. elevated plus maze, light/dark box, open field) as well as via analysis of fecal boli counts, ultrasonic vocalizations, and blood corticosterone. Male and female Sprague Dawley rats were distributed among three groups: never handled, picked-up, and handled for 5 min once daily over five days. Handled and picked-up rats spent more time in open arms and less time in closed arms of the elevated plus maze and more time in the center and less time at the perimeter of the open field compared to rats that were never handled, indicating that handled and picked-up rats were less anxious than those that were never handled. Male rats consistently defecated more frequently throughout the handling process and throughout behavioral testing, whereas females showed greater concentrations of blood corticosterone. Female rats were found to emit more 50-kHz calls and fewer 22-kHz calls compared to males. The results observed suggest that picking animals up may suffice as a handling method compared to time-intensive handling procedures, and that there are significant sex differences in response to handling.

Exposure to polystyrene microplastic beads causes sex-specific toxic effects in the model insect Drosophila melanogaster

The toxicity of MPs on aquatic creatures has been extensively studied, but little attention was paid to terrestrial organisms. To fill this gab, we conducted a series of experiments using Drosophila as a model organism to understand whether exposure to different concentrations (0.005, 0.05, 0.5 µg/ml) of polystyrene microplastics (PS-MPs) beads (2 µm in size) can impact flies feeding activity, digestion and excretion. The ability of flies to distinguish between normal and PS-MPs treated food media was tested first, and then we evaluated the effects of a 7-day short-term exposure to PS-MPs on food intake, mortality, starvation resistance, fecal pellet count, and the cellular structure of mid gut cells. The results revealed that flies can really differentiate and ignore MPs-treated food. We discovered sex-specific effects, with male flies being more sensitive to PS-MPs, with all males dying after 14 days when exposed to 0.5 µg/ml of PS-MPs, whereas female flies survived more. All male flies exposed to PS-MPs died after 24 h of starvation. Midgut cells showed concentration-dependent necrosis and apoptosis in response to PS-MPs. Our findings provide new insights into MPs toxicity on terrestrial organisms and giving a warning that management measures against MPs emission must be taken.