An Epidemiological Assessment of Cryptosporidium and Giardia spp. Infection in Pet Animals from Taiwan.

Cryptosporidium spp. and Giardia duodenalis are enteric protozoan pathogens in humans. and animals. Companion animals infected with zoonotic species/assemblages are a matter of major public concern around the world. The objectives of the present study were to determine the prevalences of Cryptosporidium spp. and G. duodenalis infections and their co-infection statuses in dogs and cats living in Taiwan and to identify the species and assemblages. Fecal samples were collected from local animal shelters (n = 285) and a veterinary hospital (n = 108). Nested polymerase chain reaction (PCR) was performed using the SSU-rRNA, ß-giardin, and glutamate dehydrogenase genes for Cryptosporidium spp. and G. duodenalis, respectively. Results showed that the overall prevalences of Cryptosporidium and G. duodenalis were 7.38% (29/393) and 10.69% (42/393). In addition, co-infection was detected in 1.02% (4/393) of all samples. Sample source, clinical sign, and breed may be risk factors that influence the infection rate. In Cryptosporidium-positive samples, C. canis and C. felis were detected most frequently. Although the canine-specific assemblages C and D (37/42) were dominant, the zoonotic human-specific assemblage A (1/42) was also found in Giardia-positive samples. Phylogenetic analysis revealed that most positive samples belonged to host-specific subtypes/assemblages, while some Cryptosporidium or Giardia-positive samples could be zoonotic. The findings suggested that pet animals could be a cause of zoonotic transmission, causing human cryptosporidiosis and giardiasis in Taiwan.

Investigation of Trehalose Supplementation Impacting Campylobacter jejuni and Clostridium perfringens from Broiler Farming.

In 2006, the European Commission banned the use of antibiotic promoters in animal feed. However, there is a new situation in poultry disease where it is necessary to study feed additives, which can overcome the diseases that were previously controlled through the addition of antibiotics and antimicrobial growth promoters in the feed. Therefore, trehalose was investigated to determine whether it impacts the growth performance and pathogenic bacteria (C. jejuni and C. perfringens) inoculation in broilers. In the first experiment, the tolerance of broilers to the addition of trehalose to their feed was investigated. There was no significant difference (p > 0.05) in body weight changes, daily weight gain, feed intake or feed conversion ratio during the feeding period. Within a 35-day feeding period, it was concluded that a trehalose dosage up to 10% does not exert a negative effect on broiler farming. Moreover, there was no significant difference (p > 0.05) in the broilers' growth performance, as well as C. jejuni and C. perfringens counts in the intestines and feces of broilers observed over a 5-week feeding period. However, Lactobacillus counts significantly increased in these groups with 3% and 5% trehalose supplementation. The findings indicate that trehalose supplementation in the feed cannot directly decrease C. jejuni and C. perfringens counts but may enhance gut health by raising Lactobacillus counts in chicken gut, particularly when enteropathogenic bacteria are present.

Are They Thinking Differently? The Perceptions and Differences in Medical Disputes between Veterinarians and Clients.

Medical disputes in veterinary practices are widespread; yet, a limited amount of research has been conducted to investigate the factors contributing to medical disputes. This study examined veterinarians' and clients' perceptions regarding risk factors and possible solutions to medical disputes. A total of 245 respondents from Taiwan, including 125 veterinarians and 120 clients, completed an electronic self-administered, semi-structured questionnaire in 2022. The questionnaire covered six dimensions: medical skills, complaint management, the attitudes of stakeholders during interactions, medical expenses, clients' perspectives, and communication modes. The results highlighted significant differences in the perceptions of risk factors for inducing medical disputes and possible solutions between clients and veterinarians in veterinary practice. First, young veterinarians and clients perceived medical skills as the highest risk factor for inducing medical disputes, while experienced veterinarians disagreed (p < 0.001). In addition, veterinarians with medical dispute experience identified stakeholders' attitudes during interactions as the top contributing factor. Second, regarding possible solutions, all veterinarians preferred offering clients cost estimates and cultivating empathy and compassion towards them. On the other hand, clients underscored the importance of obtaining informed consent for treatments and expenses and suggested that veterinarians should supply comprehensive written information to facilitate this process. This study underlies the importance of understanding stakeholders' perceptions to mitigate medical disputes and advocates for improved communication education and training for young veterinarians. These findings provide valuable insights for veterinarians and clients, contributing to preventing and managing medical disputes in veterinary practices.

Perceptions of Veterinarians and Veterinary Students on What Risk Factors Constitute Medical Disputes and Comparisons between 2014 and 2022.

This study compared the risk perceptions of medical disputes among veterinarians and veterinary students in Taiwan between 2014 and 2022. Online validity-tested questionnaires were used to collect data, with 106 (73 veterinarians and 33 students) and 157 (126 veterinarians and 31 students) surveys collected in 2014 and 2022, respectively. Respondents would be asked to rate their perceptions on how likely each risk factor constitutes a medical dispute according to their past experiences on a five-point Likert scale from 1 to 5: "Very unlikely, unlikely, neutral, likely, very likely." The results showed that overall risk perceptions increased significantly in 2022 compared to 2014, with the top risk factors being attitudes during interactions and complaint management among experienced veterinarians. In contrast, students considered medical skills and clients' perspectives as the top two risk factors, with complaints management ranking as the least significant factor. The findings suggest that effective communication and complaint management are crucial in preventing medical disputes, highlighting the importance of developing these skills in young veterinarians and veterinary students to reduce medical disputes. The study also recommends increasing practical experiences of medical disputes and complaint management in veterinary education to bridge the gap between the perceptions of experienced veterinarians and students.

Software-Based Process Simulation and Feasibility Assessment of Black Soldier Fly Larvae Fatty Acid Extraction and Fractionation.

Black soldier flies have been studied as an alternative animal feed. On the other hand, they could be used to yield an abundance of fatty acids. Their omnivorous diet and low space requirements allow for the mass breeding of black soldier flies, using widely available food wastes as feedstock. This study simulates the industrial upscaling of an extraction process for black soldier fly larvae using SuperPro™ 9.5 simulation software. This software contains an extensive material library that regulated physical data for the chemical composition of the larvae and the products. It also bundled several types of bioreactors utilized in bioprocessing. The scheduling of the plant was aided by SchedulePro, which allows for the generation of batch durations and Gantt charts. Four fatty acids were chosen as the main revenue source, with simulated proteins assigned as by-products of the plant. Ash and cellulose were the wastes of the plant, and were separated through multiple filters. The plants were later assessed for their economic feasibility. The kitchen waste plant was the most profitable, and the control variable was the only unprofitable plant. These results may have been impacted by the waste content found in the control variable and the abundance of revenue products in the kitchen-waste-fed larvae.

Bibliometric Analysis of Veterinary Communication Education Research over the Last Two Decades: Rare Yet Essential.

Research of veterinary communication education is a relatively rare but important field, and its importance has been increasingly noticed recently. This study aims to describe the existing veterinary education research literature by adopting the systematic bibliometric approach. We conducted a comprehensive literature exploration on worldwide veterinary education and veterinary communication education publications in the Web of Science Core Collection database from 1 January 2000 to 31 December 2021. VOSviewer and EXCEL were used to identify trends and patterns in characteristics of the publications, including author affiliations and countries, and the publishing journals. Based on our search criteria, in the past 22 years, there have been 6006 veterinary education publications with 101 publications in 2000, 684 publications in 2021 (577% increase), and 677 communication-related publications with 9 publications in 2000, 107 publications in 2021 (1189% increase). The VOSviewer results indicate that both the United States and England were the most vigorous countries with close collaboration. Our results show the publication quantity has been increasing at a sharp slope rate over the past twenty years, which indicates the importance and growth of veterinary education and the veterinary communication education research field, and identifies the international collaborations among countries and institutions.

Functional chicken-liver hydrolysates ameliorate insulin resistance and cognitive decline in streptozotocin-induced diabetic mice.

As part of the slaughtering processing in Taiwan, approximately 10,000 metric tons of broiler livers are produced yearly. However, these livers are regarded as waste. Our team has successfully developed a functional chicken-liver hydrolysate (CLH) with several useful activities. It has been reported that there is a positive relationship between diabetes mellitus (DM) patients and cognitive decline. To maximize broiler-livers' utilization and add value, we investigated the modulative effects of the CLHs on glucose homeostasis and cognitive decline in streptozotocin (STZ) induced diabetic mice. After a 9-wk experiment, CLH supplementation lowered blood glucose by increasing GLUT4 protein expressions in the brains, livers, and muscles of STZ-induced mice (P < 0.05). CLHs also enhanced antioxidant capacities in the livers and brains of STZ-induced mice. Amended memory and alternation behavior were tested by using water and Y-maze assays (P < 0.05). Besides, STZ-induced mice with CLH supplementation had less contracted neuron bodies in the hippocampus and lower (P < 0.05) Aß depositions in the dentate gyrus area. Less AGE accumulation and apoptosis-related proteins (RAGE, JNK, and activated Caspase 3) in the brains of STZ-induced mice were also detected by supplementing CLHs (P < 0.05). In conclusion, the results from this study offer not only scientific evidence on the amelioration of insulin resistance and cognitive decline in hyperglycemia but also add value to this byproduct.

The effects of feed supplementing Akkemansia muciniphila on incidence, severity, and gut microbiota of necrotic enteritis in chickens.

Akkermansia muciniphila (AM) is a mucin-degrading anaerobe, exerting beneficial effects on gut integrity improvement, inflammatory alleviation, and metabolic regulations in humans. Excess amounts of mucin and mucogenesis in the gut facilitate the development of necrotic enteritis (NE) in chickens. The study aimed to evaluate the effects of oral inoculation of AM on NE prevention and gut modulation in a NE-reproduced model coinfecting with Clostridium perfringens (CP) and Eimeria parasites. A total of 105 commercial 1-day-old broilers were randomly allocated into 5 groups, respectively challenged with Eimeria (Eimeria group), Eimeria and CP (Eimeria+CP group), Eimeria and CP with AM (Eimeria+CP+AM group), Eimeria and AM (Eimeria+AM group), and a placebo (Noninfected group). The treatment of AM exhibited a low degree of amelioration on NE severity. The application neither protected broilers from NE by decreasing NE-positive numbers nor reached a significant reduction in lesion scores in the small intestines. The development of NE reduced species diversity in jejunal microbiota; the pretreatments of AM exacerbated the consequence by losing species richness and promoted the similarity of the jejunal microbial community presented in the Eimeria+CP group. The participation of AM enhanced the increments of genera Clostridium sensu stricto 1 and Escherichia_Shigella and decreased the number of Lactobacillus. The significant variations of genera Clostridium sensu stricto 1 and Lactobacillus in jejunal microbiota were associated with NE development and promotion. In conclusion, oral inoculation of AM promoted the development of NE and modulated the jejunal microbiota favorable for CP overgrowth in broilers. The application of AM as a probiotic in broilers should be cautious on account of the effects to predispose NE.

Comparison of Reference-Based Assembly and De Novo Assembly for Bacterial Plasmid Reconstruction and AMR Gene Localization in Salmonella enterica Serovar Schwarzengrund Isolates.

It is well established that plasmids carrying multiple antimicrobial resistance (AMR) genes can be easily transferred among bacterial isolates by horizontal gene transfer. Previous studies have shown that a combination of short- and long-read approaches is effective in reconstructing accurate plasmids. However, high-quality Illumina short reads mapped onto the long reads in the context of an AMR hybrid monitoring strategy have not yet been explored. Hence, this study aimed to improve the reconstruction of plasmids, including the localization of AMR genes, using the above-described parameters on whole-genome sequencing (WGS) results. To the best of our knowledge, this study is the first to use S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) to confirm the number and sizes of plasmids detected by in silico-based predictions in Salmonella strains. Our results showed that de novo assembly did not detect the number of bacterial plasmids more accurately than reference-based assembly did. As this new hybrid mapping strategy surpassed de novo assembly in bacterial reconstruction, it was further used to identify the presence and genomic location of AMR genes among three Salmonella enterica serovar Schwarzengrund isolates. The AMR genes identified in the bacterial chromosome among the three Salmonella enterica serovar Schwarzengrund isolates included: AAC(3)-IV, AAC(6')-Iy, aadA2, APH(4)-Ia, cmlA1, golS, mdsA, mdsB, mdsC, mdtK, qacH, sdiA, sul2, sul3, and TEM-1 genes. Moreover, the presence of TEM-1, AAC(3)-IV, aadA2, APH(4)-Ia, cmlA1, dfrA12, floR, sul1, sul3, and tet(A) genes found within three IncFIB plasmids and one IncX1 plasmid highlight their possible transmission into the environment, which is a public health risk. In conclusion, the generated data using this new hybrid mapping strategy will contribute to the improvement of AMR monitoring and support the risk assessment of AMR dissemination.

Bacillus amyloliquefaciens and Saccharomyces cerevisiae feed supplements improve growth performance and gut mucosal architecture with modulations on cecal microbiota in red-feathered native chickens.

OBJECTIVE: The aim of study was to investigate the effects of in-feed supplementation of Bacillus amyloliquefaciens (BA) and Saccharomyces cerevisiae (SC) on growth performance, gut integrity, and microbiota modulations in red-feathered native chickens (RFCs). METHODS: A total of 18,000 RFCs in a commercial farm were evenly assigned into two dietary treatments (control diet; 0.05% BA and 0.05% SC) by randomization and raised for 11 weeks in two separate houses. Fifty RFCs in each group were randomly selected and raised in the original house with the partition for performance evaluations at the age of 9 and 11 weeks. Six non-partitioned RFCs per group were randomly selected for analyses of intestinal architecture and 16S rRNA metagenomics. RESULTS: Feeding BA and SC increased the body weight and body weight gain, significantly at the age of 11 weeks (p<0.05). The villus height/crypt ratio in the small intestines and Firmicutes to Bacteroidetes ratio were also notably increased (p<0.05). The supplementation did not disturb the microbial community structure but promote the featured microbial shifts characterized by the significant increments of Bernesiella, Prevotellaceae_NK3B31_group, and Butyrucimonas, following remarkable decrements of Bacteroides, Rikenellaceae_RC9_gut_group, and Succinatimonas in RFCs with growth benefits. Besides, functional pathways of peptidoglycan biosynthesis, nucleotide excision repair, glycolysis/gluconeogenesis, and aminoacyl transfer ribonucleic acid (tRNA) biosynthesis were significantly promoted (p<0.05). CONCLUSION: In-feed supplementation of BA and SC enhanced the growth performance, improved mucosal architectures in small intestines, and modulated the cecal microbiota and metabolic pathways in RFCs.

Comparison of Conventional Molecular and Whole-Genome Sequencing Methods for Differentiating Salmonella enterica Serovar Schwarzengrund Isolates Obtained from Food and Animal Sources.

Over the last decade, Salmonella enterica serovar Schwarzengrund has become more prevalent in Asia, Europe, and the US with the simultaneous emergence of multidrug-resistant isolates. As these pathogens are responsible for many sporadic illnesses and chronic complications, as well as outbreaks over many countries, improved surveillance is urgently needed. For 20 years, pulsed-field gel electrophoresis (PFGE) has been the gold standard for determining bacterial relatedness by targeting genome-wide restriction enzyme polymorphisms. Despite its utility, recent studies have reported that PFGE results correlate poorly with that of closely related outbreak strains and clonally dominant endemic strains. Due to these concerns, alternative amplification-based molecular methods for bacterial strain typing have been developed, including clustered regular interspaced short palindromic repeats (CRISPR) and multilocus sequence typing (MLST). Furthermore, as the cost of sequencing continues to decrease, whole genome sequencing (WGS) is poised to replace other molecular strain typing methods. In this study, we assessed the discriminatory power of PFGE, CRISPR, MLST, and WGS methods to differentiate between 23 epidemiologically unrelated S. enterica serovar Schwarzengrund isolates collected over an 18-year period from distinct locations in Taiwan. The discriminatory index (DI) of each method for different isolates was calculated, resulting in values between 0 (not discriminatory) and 1 (highly discriminatory). Our results showed that WGS has the greatest resolution (DI = 0.982) compared to PFGE (DI = 0.938), CRISPR (DI = 0.906), and MLST (DI = 0.463) methods. In conclusion, the WGS typing approach was shown to be the most sensitive for S. enterica serovar Schwarzengrund fingerprinting.

Prevalence of IncFIB Plasmids Found among Salmonella enterica Serovar Schwarzengrund Isolates from Animal Sources in Taiwan Using Whole-Genome Sequencing.

Salmonella enterica serovar Schwarzengrund is one of the most frequently isolated Salmonella serotypes responsible for human and poultry infections in Taiwan, and it has raised public health concerns. To better facilitate the understanding of transmission patterns and the dynamics of epidemics, sharing molecular data on pathogen profiles is urgently needed. The objectives of the current study were to determine and establish baseline data of S. enterica serovar Schwarzengrund isolates from 23 epidemiologically unrelated sources from year 2000 to 2018 and examine their phenotypic and genotypic characteristics. Genomic DNA of the Salmonella isolates was extracted and subjected to whole-genome sequencing using an Illumina platform. Results showed that all selected isolates exhibited multidrug resistance, and six of those were resistant to ciprofloxacin phenotypically. Genotypically, these isolates carried genes resistant to aminoglycoside (100%), phenicol (91.3%), ß-lactams (69.5%), folate pathway antagonist (100%), tetracycline (82.6%), and fluoroquinolone (4.3%). Moreover, these isolates harbor integrons with five different gene cassettes identified for the first time, which are associated with resistance to trimethoprim, streptomycin, tetracycline, sulfonamide, chloramphenicol, and gentamicin. Furthermore, prevalence of IncFIB plasmid was found among studied isolates, which may increase its ability to colonize the chicken cecum and cause extra-intestinal disease. Salmonella pathogenicity islands SPI-1 to SPI-5, SPI-13, and SPI-14, as well as C63PI locus, were also detected in all isolates. This study demonstrated that a considerable high antimicrobial resistance with high virulence levels of Salmonella were found from animal sources. Sharing data on these pathogen profiles can not only help increase the reproducibility and accessibility of genomic analysis but can also support surveillance and epidemiological investigations for salmonellosis in the region.

Ameliorative effects of functional crude-chalaza hydrolysates on the hepatosteatosis development induced by a high-fat diet.

Approximately 400 metric tons of egg chalazae, a byproduct in the liquid-egg processing plant, are produced yearly but always regarded as a waste in Taiwan. Our team successfully developed a crude egg chalaza hydrolysate by protease-A digestion (CCH-A). Free branched-chain amino acids, 3-aminoisobutyric acid, and ß-alanine, and anserine were assayed in the CCH-A used in this study. Besides, the in vitro bile-acid binding ability and inhibitory lipase activity of CCH-As were demonstrated. Then, high-fat diet feeding for 10 wk caused hyperlipidemia, insulin resistance, and hepatosteatosis in hamsters (P < 0.05). However, CCH-A co-treatment decreased serum/liver triglyceride levels and lipid accumulation in livers by increasing daily fecal lipid/bile-acid outputs, upregulating fatty-acid ß oxidation, and downregulating fatty-acid biosynthesis in livers (P < 0.05). CCH-A co-treatment also amended insulin resistance, augmented hepatic antioxidant capacity, and decreased liver damages and inflammatory responses (P < 0.05). Taken together, our results do not only demonstrate the hepatoprotective effects of CCH-As against a chronic high-fat dietary habit, achieving effects similar to Simvastatin, but also decrease the environmental burden of handling chalazae in the liquid-egg industry.

Coexistence of Fosfomycin Resistance Determinant fosA and fosA3 in Enterobacter cloacae Isolated from Pets with Urinary Tract Infection in Taiwan.

To analyze the characteristics of fosA and fosA3 in Enterobacter cloacae isolated from aspirated and catheterized urine culture specimens of companion pets in Taiwan. A total of 19 E. cloacae isolates from pets with urinary tract infection were screened for the presence of fosA, fosA3, and fosC2 and for the genetic context of them by PCR amplification and sequencing. The transferability, resistance phenotypes, plasmid replicon typing properties and genetic environments of fosA- and/or fosA3-positive strains were characterized. Five E. cloacae isolates were positive for fosA and three coharbored fosA and fosA3. No fosC determinant was detected. Transconjugants of fosA3 were successfully acquired, while the acquisition of fosA transconjugants was failed. The minimum inhibitory concentrations (MICs) of the three fosA3-positive isolates and their transconjugants were ≥256 mg/L, whereas the MICs of the five fosA-positive isolates ranged from 64 mg/L to 256 mg/L. Three plasmid replicons (InCFrepB, InCL/M, and InCHI2) were identified in fosA- and fosA3-positive E. cloacae isolates. Different genetic contexts lay in the downstream region of fosA and fosA3, respectively. Eight distinct patterns based on the similarity value of more than 80% were typed for all the 8 fosA-positive isolates. In conclusion, the fosA concomitant with fosA3 were found in E. cloacae isolates. The fosA3 not only exhibits stronger activity of inactivating fosfomycin than fosA but also possesses stronger potential to spread than fosA. Different genetic backgrounds exist in these fosA- and fosA3-positive isolates, and different mobile elements may confer the dissemination of fosA and fosA3.

Genome-wide DNA methylation analysis using MethylCap-seq in canine high-grade B-cell lymphoma.

DNA methylation is a comprehensively studied epigenetic modification and plays crucial roles in cancer development. In the present study, MethylCap-seq was used to characterize the genome-wide DNA methylation patterns in canine high-grade B-cell lymphoma (cHGBL). Canine methylated DNA fragments were captured and the MEDIUM-HIGH and LOW fraction of methylated DNA was obtained based on variation in CpG methylation density. In the MEDIUM-HIGH and LOW fraction, 2144 and 1987 cHGBL-specific hypermethylated genes, respectively, were identified. Functional analysis highlighted pathways strongly related to oncogenesis. The relevant signaling pathways associated with neuronal system were also revealed, echoing recent novel findings that neurogenesis plays key roles in tumor establishment. In addition, 14 genes were hypermethylated in all the cHGBL cases but not in the healthy dogs. These genes might be potential signatures for tracing cHGBL, and some of them have been reported to play roles in various types of cancers. Further, the distinct methylation pattern of cHGBL showed a concordance with the clinical outcome, suggesting that aberrant epigenetic changes may influence tumor behavior. In summary, our study characterized genome-wide DNA methylation patterns using MethylCap-seq in cHGBL; the findings suggest that specific DNA hypermethylation holds promise for dissecting tumorigenesis and uncovering biomarkers for monitoring the progression of cHGBL.

Modulation effects of black-vinegar-based supplement against a high-fat dietary habit: antiobesity/hypolipidemic, antioxidative, and energy-metabolism effects.

BACKGROUND: An imbalanced fat or excess energy intake always results in obesity and increased serum/liver lipids, thus leading to metabolic syndromes. Given the bioactive components in black vinegar (BV), such as branched amino acids, phenolic profile, and mineral contents, we investigated the antiobesity effects of BV-based supplements in rats fed a high-fat diet (HFD). RESULTS: HFD (30% fat, w/w) feeding increased (P < 0.05) body weight, weight gains, weights of livers and mesenteric, epididymal, and perirenal adipose tissues, and serum/liver triglyceride levels relative to those of rats fed a normal diet (4% fat, w/w; CON). These increased values were ameliorated (P < 0.05) by supplementing with BV-based supplements but were still higher (P < 0.05) than those of CON rats. The increased areas of perirenal adipocytes in rats fed with an HFD were also decreased (P < 0.05) by supplementing with BV-based supplements, which might result from an upregulation (P < 0.05) of 5'-adenosine monophosphate-activated protein kinase (AMPK), carnitine palmitoyltransferase-1 (CPT1), and uncoupling protein-2 (UCP2) in the perirenal adipose tissues. A similar effect was observed for AMPK, peroxisome proliferator-activated receptor alpha, retinoid X receptor alpha, CPT1, and UCP2 gene and protein levels in livers (P < 0.05). Generally, BV-based supplements increased the fecal triglyceride, cholesterol, and bile acid levels of rats fed with an HFD, which partially contribute to the lipid-lowering effects. Furthermore, BV-based supplements increased (P < 0.05) hepatic Trolox equivalent antioxidant capacity and lowered (P < 0.05) serum/liver thiobarbituric acid reactive substances values in HFD-fed rats. CONCLUSION: In a chronic high-fat dietary habit, the food-grade BV-based supplement is a good daily choice to ameliorate obesity and its associated comorbidities. © 2020 Society of Chemical Industry.

Analysis of steroid hormones in shell eggs from layer breeds common to Taiwan by liquid chromatography-tandem mass spectrometry.

Steroid hormones are often used in animal agriculture but are currently banned for use in domesticated fowl because residual hormones could be present in eggs for human consumption. Egg samples from eight common commercial poultry layer breeds (Hy-Line W-36, Hy-Line Brown, ISA-White, ISA-Brown, Lohnmann Ultra-Lite, Lohnmann-Brown, Hisex White, Hisex Brown) in Taiwan were screened for a combination of 15 natural and synthetic steroid hormones by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for consumer assurance. Only natural hormones such as progesterone, 4-androstene-3,17-dione, and testosterone were detected. Regarding each breed, the interaction effect (age × shell color), main effect (age or shell color), and blocking effect (lighting system) were further analyzed by using 2 × 2 factorial arrangement of treatment in a randomized block design. We also discovered associations between yolk steroid hormone levels and laying hen age, as well as lighting conditions. Additionally, we found a correlation between hormone levels and eggshell color, suggesting a potential role in brown pigmentation. Ultimately, we concluded that detectable steroid hormone levels in eggs were not a consumer health risk. Furthermore, these data provide empirical hormone concentrations in various types of commercial layer breeds for future research.

The netB-positive Clostridium perfringens in the experimental induction of necrotic enteritis with or without predisposing factors.

The netB-positive Clostridium perfringens has been considered as the requisite to consistently induce necrotic enteritis (NE). However, use of a netB-positive strain did not guarantee consistent NE reproduction unless high protein diets or Eimeria, conceived as 2 major predisposing factors, was incorporated. To establish a refined model, the roles of dietary fishmeal inclusion, Eimeria inoculation, and netB-positive C. perfringens challenge in NE induction and the confounding effects of Eimeria infection on NE were examined. The results showed that the use of netB-positive C. perfringens without a predisposing factor failed to induce NE. Fishmeal incorporation promoted the occurrence of NE but did not significantly affect the incidence of the disease in conjunction with challenge of netB-positive C. perfringens. However, the additional participation of Eimeria infection in the same induction procedure produced significantly higher numbers of NE cases and promoted more severe lesions in chickens (P < 0.05). Inoculation of Eimeria resulted in a significant higher incidence of NE compared to the non-Eimeria treated group (P < 0.05). The results demonstrated that both netB-positive C. perfringens and predisposing factors were required for the reproduction of disease. Mild-to-moderate coccidial infection (coccidial lesion score ≤ 2) was noted in NE cases in this model but severe coccidial infection did not correlate with the occurrence of NE, indicating mild coccidial infection may be beneficial for the development of NE. If multiple species infection of Eimeria precedes the challenge of C. perfringens, days 19 to 21 (1 to 3 D after the last clostridial challenge) was the time period favorable for observations of NE lesions. The time after this period may be subject to bias of severity, incidence, or mortality of NE owing to the profound coccidial lesions in the intestinal region. This study demonstrated that the co-infection with netB-positive C. perfringens and Eimeria species under fishmeal incorporation produced a desirable NE model, being of value in studying the effectiveness of novel feed additives and alternative mitigation strategies to prevent NE.

Characterization of Escherichia coli Isolated from Day-old Chicken Fluff in Taiwanese Hatcheries.

Avian colibacillosis resulting from avian pathogenic Escherichia coli (APEC) seriously disrupts poultry production. Hatcheries are the main source of chickens for commercial farms. To characterize the potential pathogenicity of E. coli strains isolated from hatcheries, 2344 fluff samples from 1-day-old chickens were collected from hatching incubators between October 2016 and November 2017. Among the hatcheries, the incidence of E. coli varied from 0% to 16.9%, with an overall incidence of 2.0%. High incidences reflected inadequate sanitation in some hatcheries. We also compared 20 clinically isolated APEC strains with fluff-originated E. coli in terms of existence of 10 virulence-associated genes (VAGs) and antimicrobial-resistance genes, and antimicrobial resistance using minimum inhibitory concentration (MIC) values. Our results showed that APEC more-frequently possessed most of the assessed VAGs (papC, astA, cvaC, hlyF, fyuA, iroN, iutA, iss, and ompT), suggesting that fluff-originated E. coli is less likely to cause avian colibacillosis. However, fluff-originated E. coli more-frequently expressed the adhesion gene fimC, which could confer higher upper respiratory tract adhesion. Both APEC and fluff-originated E. coli demonstrated multidrug resistance including 100% resistance to ampicillin, amoxicillin, cephalexin, florfenicol, and trimethoprim-sulfamethoxazole. Based on median MIC values, fluff-originated E. coli was more susceptible to antibiotics. However, resistance-gene existence did not significantly differ between groups, suggesting that fluff-originated E. coli should still be a public health concern. Molecular subtyping with XbaI-digested pulsed-field gel electrophoresis revealed that only a few strains showed identical patterns, indicating that a variety of contamination sources were present within individual hatcheries. Identical strains within the same hatchery may indicate vertical transmission from parent flocks. Overall, this is the first study to characterize fluff-originated E. coli. Our results suggest that it has lower pathogenicity than APEC and that thorough sanitation should be performed to reduce the occurrence of fluff-originated E. coli in hatcheries.

Caracterización de Escherichia coli aislada de plumón de pollitos de un día en plantas de incubación en Taiwán. La colibacilosis aviar resultante de Escherichia coli patogénica aviar (APEC) puede afectar de manera severa a la producción avícola. Las plantas de incubación son la fuente principal de pollos para las granjas comerciales. Para caracterizar la patogenicidad potencial de las cepas de E. coli aisladas de las plantas de incubación, se recolectaron 2344 muestras de plumón de pollos de un día de edad de plantas de incubación entre octubre del 2016 a noviembre del 2017. Entre las plantas de incubación, la incidencia de E. coli fue del cero por ciento hasta el 16.9%, con una incidencia global del 2.0%. Las altas incidencias reflejaron una sanidad inadecuada en algunas plantas. También se compararon 20 cepas de E. coli patogénica aviar aisladas de casos clínicos con E. coli originada del plumón con relación a la presencia de 10 genes asociados a la virulencia (VAG), con genes asociados con la resistencia a los antimicrobianos y se evaluó la resistencia a los antimicrobianos mediante valores de concentración mínima inhibitoria (MIC). Los resultados mostraron que la E. coli patogénica aviar poseía con mayor frecuencia la mayoría de los genes asociados con virulencia evaluados (papC, astA, cvaC, hlyF, fyuA, iroN, iutA, iss y ompT), lo que sugiere que la E. coli originada del plumón es menos probable que cause colibacilosis aviar. Sin embargo, la E. coli originada de plumón expresó con mayor frecuencia el gene de adhesión fimC, que podría conferir una mayor adhesión en el tracto respiratorio superior. Tanto la E. coli patogénica aviar como la E. coli originada del plumón demostraron resistencia a múltiples fármacos, incluida una resistencia del 100% a la ampicilina, amoxicilina, cefalexina, florfenicol y trimetoprimsulfametoxazol. Con base la mediana de los valores de la concentración mínima inhibitoria, la E. coli originada del plumón fue más susceptible a los antibióticos. Sin embargo, la existencia del gene de resistencia no fue diferente significativamente entre los grupos, lo que sugiere que la E. coli originada del plumón puede ser un problema de salud pública. El subtipo molecular con electroforesis en gel de campo pulsado digerido con XbaI reveló que solo unas pocas cepas mostraron patrones idénticos, lo que indica que una variedad de fuentes de contaminación estaban presentes en las plantas de incubación individuales. Las cepas idénticas dentro de la misma planta de incubación pueden indicar transmisión vertical a partir de las parvadas paternas. En general, este es el primer estudio en caracterizar E. coli originada del plumón. Estos resultados sugieren que tiene una patogenicidad más baja que la E. coli patogénica aviar y que se debe realizar una limpieza y desinfección exhaustivas para reducir la aparición de E. coli originada de plumón en las plantas de incubación.

Analysis of gut microbiota and the effect of lauric acid against necrotic enteritis in Clostridium perfringens and Eimeria side-by-side challenge model.

Gut microbiota has been demonstrated to be involved in intestinal nutrition, defense, and immunity, as well as participating in disease progression. This study was to investigate gut microbiota changes in chickens challenged with netB-positive Clostridium perfringens strain (CP1) and/or the predisposing Eimeria species (Eimeria) and fed diets with fishmeal supplementation. In addition, the effects of lauric acid, a medium-chain fatty acid (MCFA), on necrotic enteritis (NE) reduction and modulation of microbiota were evaluated. The results demonstrated that microbial communities in the jejunum were distinct from those in the cecum, and the microbial community change was more significant in jejunum. Challenge of CP1 in conjunction with Eimeria significantly reduced species diversity in jejunal microbiota, but cecal microbiota remained stable. In the jejunum, CP1 challenge increased the abundance of the genera of Clostridium sensu stricto 1, Escherichia Shigella, and Weissella, but significantly decreased the population of Lactobacillus. Eimeria infection on its own was unable to promote NE, demonstrating decrements of Clostridium sensu stricto 1 and Lactobacillus. Co-infection with CP1 and Eimeria reproduced the majority of NE lesions with significant increment of Clostridium sensu stricto 1 and reduction in Lactobacillus. The advance of changes on these two taxa increased the severity of NE lesions. Further analyses of metagenomeSeq, STAMP, and LEfSe consistently showed significant overgrowth of Clostridium sensu stricto 1 was associated with NE. The supplementation of lauric acid did not reduce NE incidence and severity but decreased the relative abundance of Escherichia Shigella. In conclusion, significant overgrowth of C. perfringens as well as other Clostridium species in Clostridium sensu stricto 1 with the decrement of Lactobacillus in the jejunum is the featured microbiota correlated with NE. Controlling proliferation of Clostridium sensu stricto 1 and manipulation of Lactobacillus in the jejunum should be the strategy to prevent NE.