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Helminth infection is highly prevalent in indigenous chickens reared in semi-scavenging/ scavenging systems in Bangladesh. Here, we estimated the prevalence of gizzard worm infection in indigenous chickens, the detection of the worm-induced pathologies, the development of ex vivo cultural protocol, and anthelmintic efficacy. We randomly collected and examined 390 chickens and isolated worms from the gizzard and proventriculus. The isolated worms were identified as Cheilospirura hamulosa Diesing, 1861. The overall prevalence of C. hamulosa was 33.1% (129 out of 390). Prevalence of the worm was almost similar in both sexes but significantly (p <0.05) higher in adult chickens (44.3%) and in the summer season (47.1%). In heavy infections, C. hamulosa destroyed the muscular layer of the gizzard. The presence of brown necrotic tissues and curd-like caseous materials was detected in the affected gizzards. In severe cases, the horny lining of the gizzard was inflamed, necrotized and marked by multiple holes and brick-red colored spots. Liquefied, fetid materials oozed out from the muscular layer in extensive cases. Histopathological examination showed marked infiltrations of eosinophils. In serum-supplemented M199 and DMEM, adult C. hamulosa survived well and reproduced. Levamisole (LEV) and ivermectin (IVM) efficiently killed the worm. However, albendazole (ABZ), mebendazole (MBZ) and piperazine (PPZ) did not kill the worms. Our results suggest that C. hamulosa is highly prevalent in semi-scavenging chickens in Bangladesh. LEV and IVM can be used to treat and control the infection in chickens.

While mechanical properties of the brain have been investigated thoroughly, the mechanical properties of human brain tumors rarely have been directly quantified due to the complexities of acquiring human tissue. Quantifying the mechanical properties of brain tumors is a necessary prerequisite, though, to identify appropriate materials for surgical tool testing and to define target parameters for cell biology and tissue engineering applications. Since characterization methods vary widely for soft biological and synthetic materials, here, we have developed a characterization method compatible with abnormally shaped human brain tumors, mouse tumors, animal tissue and common hydrogels, which enables direct comparison among samples. Samples were tested using a custom-built millimeter-scale indenter, and resulting force-displacement data is analyzed to quantify the steady-state modulus of each sample. We have directly quantified the quasi-static mechanical properties of human brain tumors with effective moduli ranging from 0.17-16.06 kPa for various pathologies. Of the readily available and inexpensive animal tissues tested, chicken liver (steady-state modulus 0.44 ± 0.13 kPa) has similar mechanical properties to normal human brain tissue while chicken crassus gizzard muscle (steady-state modulus 3.00 ± 0.65 kPa) has similar mechanical properties to human brain tumors. Other materials frequently used to mimic brain tissue in mechanical tests, like ballistic gel and chicken breast, were found to be significantly stiffer than both normal and diseased brain tissue. We have directly compared quasi-static properties of brain tissue, brain tumors, and common mechanical surrogates, though additional tests would be required to determine more complex constitutive models.

Adenoviral gizzard erosion is an emerging disease with negative impact on health and production of chickens. In this study, we compared in vitro and in vivo characteristics of a fowl adenovirus serotype 1 (FAdV-1), attenuated by 53 consecutive passages in primary chicken embryo liver (CEL) cell cultures (11/7127-AT), with the virulent strain (11/7127-VT). Whole genome analysis revealed near-complete sequence identity between the strains. However, a length polymorphism in a non-coding adenine repeat sequence (11/7127-AT: 11 instead of 9) immediately downstream of the hexon open reading frame was revealed. One-step growth kinetics showed delayed multiplication of 11/7127-AT together with significantly lower titers in cell culture (up to 4 log10 difference), indicating reduced replication efficiency in vitro. In vivo pathogenicity and immunogenicity were determined in day-old specific pathogen-free layer chicks inoculated orally with the respective viruses. In contrast to birds infected with 11/7127-VT, birds infected with 11/7127-AT did not exhibit body weight loss or severe pathological lesions in the gizzard. Virus detection rates, viral load in organs and virus excretion were significantly lower in birds inoculated with 11/7127-AT. Throughout the experimental period, these birds did not develop measurable neutralizing antibodies, prevalent in birds in response to 11/7127-VT infection. Differences in pathogenicity between the virulent FAdV-1 and the attenuated strain could not be correlated to prominently discriminate genomic features. We conclude that differential in vitro growth profiles indicate that attenuation is linked to modulation of viral replication during interaction of the virus with the host cells. Thus, hosts would be unable to prevent the rapid replication of virulent FAdV leading to severe tissue damage, a phenomenon broadly applicable to further FAdV serotypes, considering the substantial intra-serotype virulence differences of FAdVs and the variation of diseases.

An experiment was carried out to study the effect of different forms of wheat (airtight silo stored whole wheat, conventionally stored whole wheat, and ground wheat included in pellets) and dietary xylanase addition on production results and gastrointestinal characteristics of broiler chickens. Ileal viscosity, pancreatic digestive enzyme activities, and the composition and activity of the intestinal microflora were considered as response parameters. Differences between the 2 types of whole wheat with respect to the various measured parameters were marginal, whereas distinct differences were found between pellet-fed birds and birds receiving whole wheat. Whole wheat feeding improved feed conversion ratio and reduced water consumption (P < 0.001). Compared with pellets, whole wheat increased the relative weight of pancreas and gizzard and the dry matter concentration of gizzard content (P < 0.001). Whole wheat feeding reduced the pH in the gizzard contents (P < 0.01) and increased ileal viscosity. The addition of xylanase reduced ileal viscosity in birds receiving whole wheat to the same level as in pellet-fed birds. Whole wheat feeding resulted in lower activities of amylase in pancreatic tissue (P = 0.054), whereas xylanase addition increased chymotrypsin (P = 0.030) and lipase activities (P = 0.052). Whole wheat feeding resulted in lower intestinal numbers of lactose-negative enterobacteria (P < 0.05) and tended to reduce the ileal and cecal numbers of Clostridium perfringens (P less than or equal to 0.08). It is concluded that whole wheat feeding stimulates gizzard function, which in turn prevents potentially pathogenic bacteria from entering the intestinal tract.

Arsenic (As) and copper (Cu) are ubiquitous pollutants that pose a threat to the environment. Our aim is to study the underlying mechanisms by which As and Cu act on the chicken gizzard. In order to detect ionic disorders in chicken gizzard under chronic treatment with As 3+ and/or Cu 2+ and whether they can induce oxidative damage as well as immune disorders, 30 mg/kg arsenic trioxide (As 2 O 3 ) and/or 300 mg/kg copper sulfate (CuSO 4 ) were added to the chicken’s basal diet. After 12 weeks of exposure, trace elements were found to have significant interference, accompanied by damage to the antioxidant system. In addition, As 3+ and/or Cu 2+ activated the nuclear factor kappa B (NF-κB), inducing severe inflammation. At the same time, damaged structural integrity which might be caused by inflammation was discovered after hematoxylin and eosin (H&E) staining. Moreover, symbolic Th1/Th2 (Th, helper T cell) drift was also observed in treatment groups, meaning that immune function is left to be affected, and the increment in heat shock proteins may be a self-protective mechanism of gizzard. Interestingly, we found that the damage to the gizzard of chicken was aggravated in a time-dependent manner, and the combined exposure was more pathogenic than the single exposure, of which the mechanism needs further exploration. Together, this work helps move us toward a better understanding of the molecular mechanisms that mediate the interactions between Cu excess and As 3+ exposures and possible health consequences in susceptible species.

Age-related increases in the repeatable expression of labile phenotypic traits are often assumed to arise from an increase in among-individual variance due to differences in developmental plasticity or by means of state-behavior feedbacks. However, age-related increases in repeatability could also arise from a decrease in within-individual variance as a result of stabilizing trait expression, that is, canalization. Here we describe age-related changes in within-individual and among-individual variance components in two correlated traits—gizzard mass and exploration behavior—in a medium-sized shorebird, the red knot (Calidris canutus). Increased repeatability of gizzard mass came about due to an increase in among-individual variance, unrelated to differences in developmental plasticity, together with decreases in within-individual variance consistent with canalization. We also found canalization of exploration but no age-related increase in overall repeatability, which suggests that showing predictable expression of exploration behavior may be advantageous from a very young age onward. Contrasts between juveniles and adults in the first year after their capture provide support for the idea that environmental conditions play a key role in generating among-individual variation in both gizzard mass and exploration behavior. Our study shows that stabilization of traits occurs under constant conditions: with increased exposure to predictable cues, individuals may become more certain in their assessment of the environment allowing traits to become canalized.

Although plastic pollution happens globally, the micro- (<5 mm) and macroplastic (5–150 mm) transfer of plastic to terrestrial species relevant to human consumption has not been examined. We provide first-time evidence for micro- and macroplastic transfer from soil to chickens in traditional Mayan home gardens in Southeast Mexico where waste mismanagement is common. We assessed micro- and macroplastic in soil, earthworm casts, chicken feces, crops and gizzards (used for human consumption). Microplastic concentrations increased from soil (0.87 ± 1.9 particles g −1 ), to earthworm casts (14.8 ± 28.8 particles g −1 ), to chicken feces (129.8 ± 82.3 particles g −1 ). Chicken gizzards contained 10.2 ± 13.8 microplastic particles, while no microplastic was found in crops. An average of 45.82 ± 42.6 macroplastic particles were found per gizzard and 11 ± 15.3 macroplastic particles per crop, with 1–10 mm particles being significantly more abundant per gizzard (31.8 ± 27.27 particles) compared to the crop (1 ± 2.2 particles). The data show that micro- and macroplastic are capable of entering terrestrial food webs.

Recently, outbreaks of adenoviral gizzard erosion (AGE) have been documented in pullets and layers housed free range and in enriched cage systems characterized by increased mortality and a negative impact on egg production. In the present study the pathogenicity of a fowl adenovirus serotype 1 (FAdV-1) field strain as well as the aetiological role of a FAdV-8a strain, both isolated from AGE affected pullets, were investigated in vivo in 20-week-old specific-pathogen-free (SPF) layer-type chickens. Furthermore, the efficacy of a single (week 17) and double (week 14 and 17) application of a live vaccine consisting of an apathogenic FAdV-1 (CELO strain) against challenge with virulent FAdV-1 was investigated. For the first time, AGE was successfully reproduced in adult birds after oral infection of 20-week-old SPF birds with a virulent FAdV-1 field isolate, characterized by pathological changes of the gizzard from 7 days post challenge onwards. In addition, a negative impact of the FAdV-1 infection on the development of the reproductive tract was observed. Thus, confirming the pathogenicity and aetiological role of FAdV-1 in the development of AGE and economic losses due to AGE in layers. In contrast, no pathological changes were observed in birds infected with FAdV-8a. Independent of a single or double application of the live FAdV-1 vaccine strain CELO, no gross pathological changes were observed in gizzards post challenge with the virulent FAdV-1, indicating that complete protection of layers against horizontal induction of AGE was achieved. Nonetheless, virulent FAdV-1 was detected in cloacal swabs and gizzards in both vaccinated groups post challenge determined by the application of an amplification refractory mutation system quantitative PCR used to differentiate between vaccine and challenge strains.

Avian influenza virus subtype H9N2 is identified in chickens with respiratory disease while Bacillus cereus ( B. cereus ) has been frequently isolated from chicken feed in China. However, the roles of co-infection with these two pathogens remain unclear. In the present study, SPF chicks were intragastrically administered with 10 8  CFU/mL of B. cereus for 7 days and then inoculated intranasally with 100 EID 50 of H9N2 three days later. Alternatively, chickens were initially inoculated with H9N2 and then with B. cereus for one week. Post administration, typical respiratory distress persisted for 5 days in both co-infection groups. Gizzard erosions developed in the groups B. cereus /H9N2 and B. cereus group on 7 th day while in group H9N2/ B. cereus on 14 th day. More importantly, both air-sac lesions and lung damage increased significantly in the co-infection group. Significant inflammatory changes were observed in the B. cereus group from day 7 to day 21. Moreover, higher loads of H9N2 virus were found in the co-infected groups than in the H9N2 group. Newcastle Disease Virus (NDV) specific antibodies were decreased significantly in the H9N2/ B. cereus group compared to the B. cereus and the B. cereus /H9N2 groups. Nonspecific IgA titers were reduced significantly in the B. cereus group and the H9N2/ B. cereus group compared to the control group. In addition to this, lower lymphocyte proliferation was found in the con-infection groups and the H9N2 group. Hence, feed-borne B. cereus contamination potentially exacerbates gizzard ulceration and aggravates H9N2-induced respiratory distress by inhibiting antibody-mediated immunity and pathogen clearance. Thus controlling the B. cereus contamination in poultry feed is immediately needed.

Alteration of sphingolipid synthesis is a key event in fumonisins toxicity, but only limited data have been reported regarding the effects of fumonisins on the sphingolipidome. Recent studies in chickens found that the changes in sphingolipids in liver, kidney, lung, and brain differed greatly. This study aimed to determine the effects of fumonisins on sphingolipids in heart, gizzard, and breast muscle in chickens fed 20.8 mg FB1 + FB2/kg for 9 days. A significant increase in the sphinganine:sphingosine ratio due to an increase in sphinganine was observed in heart and gizzard. Dihydroceramides and ceramides increased in the hearts of chickens fed fumonisins, but decreased in the gizzard. The dihydrosphingomyelin, sphingomyelin, and glycosylceramide concentrations paralleled those of ceramides, although the effects were less pronounced. In the heart, sphingolipids with fatty acid chain lengths of 20 to 26 carbons were more affected than those with 14–16 carbons; this difference was not observed in the gizzard. Partial least squares-discriminant analysis on sphingolipids in the heart allowed chickens to be divided into two distinct groups according to their diet. The same was the case for the gizzard. Pearson coefficients of correlation among all the sphingolipids assayed revealed strong positive correlations in the hearts of chickens fed fumonisins compared to chickens fed a control diet, as well as compared to gizzard, irrespective of the diet fed. By contrast, no effect of fumonisins was observed on sphingolipids in breast muscle.