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Effects of transient prepubertal 6-N-propyl-2-thiouracil treatment on testis development and function in the domestic fowl.

It has been well established that thyroid hormones play an important role in regulating the onset of puberty and reproductive function in birds. In mammals it has been shown that transient hypothyroidism induced with the reversible goitrogen 6-N-propyl-2-thiouracil (PTU) can result in tremendous increases in testis size and sperm production and that the timing of hypothyroidism must correspond to the period of Sertoli cell proliferation. As the period of Sertoli cell proliferation is not precisely known in the fowl, an experiment was conducted to determine whether chicken testes have a similar window of sensitivity to PTU treatment. Broiler breeder male chicks (Peterson) were placed in floor pens at one day of age and reared according to the breeder's management guide for the entire 28-wk duration (controls) or up to the point of dietary treatment with PTU (0.1% w:w) for 6 wk that began at 2-wk intervals (2-8, 4-10, 6-12, 8-14, and 10-16 wk of age); after treatment, birds were returned to feed restriction and photostimulated at 20 wk of age. Birds were bled and killed, and testes were collected at 4-wk intervals. At 28 wk, one testis was fixed for histological examination and one was immediately placed in liquid N2 for sperm counts. Treatment with PTU from 6 to 12 wk of age resulted in a 96% increase in mean testis weight at 28 wk of age (treated 39.3 +/- 4.1 g per testis vs. control 20.0 +/- 1.6 g per testis). These testes exhibited normal morphology and increased relative sperm production. Treatment with PTU from either 8 to 14 or 10 to 16 wk of age resulted in approximately a 35% increase in testis mass at 28 wk of age relative to the control value (27.2 +/- 2.0 g and 27.7 +/- 3.6 g vs. 20.0 +/- 1.6 g per testis, respectively). However, both of these groups clearly demonstrated precocious puberty and abnormal spermatogenesis. These results suggest that appropriately timed PTU treatment may result in permanent increases in testis size and sperm production in the domestic fowl.

Animals↗

Sperm mobility: A primary determinant of fertility in the domestic fowl (Gallus domesticus).

Previous research demonstrated that sperm mobility is a quantitative trait of the domestic fowl. The trait is quantified by measuring the absorbance of an Accudenz solution after overlay with a sperm suspension and brief incubation at body temperature. In the present work, average and high sperm mobility phenotypes (n = 30 males per phenotype) were selected from a base population. Differences were found between sperm oxygen consumption (p < 0.0001), acylcarnitine content (p < 0.05), linear velocity (p < 0.001), and straightness (p < 0.001), a trajectory variable measured with the Hobson SpermTracker. Oxygen consumption and stearoylcarnitine content of sperm from the high-mobility phenotype were twice those observed with sperm from average males, implying a pivotal role for mitochondria. On the basis of these results, a graded relationship was predicted between fertility and sperm mobility. Males (n = 48) were chosen at random from another base population, sperm mobility was measured per male, and each ejaculate was used to inseminate 8-12 hens (8 x 10(7) viable sperm per hen). When fertility was plotted as a function of sperm mobility, data points approximated a skewed logistic function. The hypothesis that vaginal immunoglobulins constitute an immunological barrier to sperm transport was tested and rejected. Therefore, we concluded that sperm mobility is a primary determinant of fertility in the fowl.

Animals↗

Indirect partner choice through manipulation of male behaviour by female fowl, Gallus gallus domesticus.

The direct and indirect consequences of female copulatory behaviour for copulation success have seldom been quantified. In feral fowl, most copulations were forced by males and copulation success was determined by two factors. First, female differential resistance and solicitation directly affected copulation success and were displayed non-randomly with respect to male social status. Second, another female copulatory behaviour, the distress call, had an indirect effect on both copulation success and the quality of copulation partners. Distress calls triggered male attention to a copulation, which increased the probability of higher-ranking males than the copulating male disrupting the copulation and inseminating the calling female. Females preferentially uttered distress calls when mounted by low-ranking males. Both copulation resistance and distress calling influenced copulation success, but only distress calling increased the probability of copulation disruption by other males. Consistent with the effect of direct selection, differential distress calling indirectly biased copulation success in favour of dominant males. Female fowl may thus ameliorate the effect of male sexual coercion by manipulating male behaviour.

Animals↗

Evidence for the involvement of influenza A (fowl plague Rostock) virus protein P2 in ApG and mRNA primed in vitro RNA synthesis.

Eleven temperature-sensitive (ts) mutants of influenza A (fowl plague, Rostock) virus were analysed for in vitro RNA transcriptase activity in reactions primed by ApG or globin mRNA at 31 degrees C or at 40.5 degrees C, the restrictive temperature for ts mutant growth. Only those ts mutants studied which were defective in RNA segment 1, coding for the virion P2 protein, were defective in RNA transcriptase activity when compared to wild-type virus. Mutants having a defect in the P2 protein had no significant RNA transcriptase activity in reactions at 40.5 degrees C primed by globin mRNA. However, one mutant showed RNA transcriptase activity similar to wild-type virus at 40.5 degrees C when ApG (0.3 mM) was used as primer. The results suggest that influenza (fowl plague, Rostock) P2 protein is directly involved in the mRNA priming reaction, as well as in the RNA transcription reaction in vitro.

Adenosine Monophosphate↗

Genetic diversity of Pasteurella multocida fowl cholera isolates as demonstrated by ribotyping and 16S rRNA and partial atpD sequence comparisons.

The genetic diversity of Pasteurella multocida, the aetiological agent of fowl cholera, was investigated. The strain collection comprised 69 clinical isolates representing a wide spectrum of hosts and geographic origin. The three type strains for the subspecies of P. multocida were also included. Avian isolates of P. multocida subsp. multocida and P. multocida subsp. septica did not represent separate lines by HpaII ribotyping and the two type strains of mammalian origin (porcine and cat bite) seemed to be representative of avian strains of P. multocida subspp. multocida and septica. By ribotyping, all P. multocida subsp. gallicida strains, except one chicken isolate and the type strain, clustered together. This indicated that the bovine type strain was not representative of this subspecies and that most strains of P. multocida subsp. gallicida are genetically related and may be distantly related to other P. multocida isolates, including those of avian origin. By 16S rRNA and atpD sequence comparisons of selected strains, including both P. multocida isolated from birds and mammals and selected distantly related Pasteurella species associated with birds and mammals, it was found that P. multocida is monophyletic. Extended DNA-DNA hybridizations are highly indicated since strains may exist which would connect the existing subspecies at species level. The considerable genetic diversity of P. multocida fowl cholera isolates is probably related to the clonal nature of this organism, resulting in many divergent lines.

Animals↗

Acquired immunity to larvae of Amblyomma marmoreum and A.hebraeum by tortoises, guinea-pigs and guinea-fowl.

A study of acquired resistance in guinea-pigs, guinea-fowl and tortoises to larvae of the ticks Amblyomma hebraeum Koch and A.marmoreum Koch (Acari: Ixodidae) showed that repeated infestations of the laboratory host resulted in a 29.3-49.4% reduction respectively in weight of engorged larvae. No resistance was induced in guinea-fowl, the natural host for larvae of both species. A small decline in the mean weight of engorged larvae was observed for both species of ticks after three successive infestations on tortoises. The reduction in weight was more pronounced in A.hebraeum (14.5%) which does not naturally feed on tortoises than for A.marmoreum (8.4%), a common tick on this host. The occurrence of acquired resistance in natural tick host relationships as opposed to unnatural tick host relationships is discussed.

Animals↗

Amino-acid sequence of feather keratin from fowl.

The amino acid sequence of a single polypeptide chain, B-4, from fowl feather barbs has been determined. The B-4 chain was found to consist of 96 amino acid residues and to have a molecular weight of 10206 in the S-carboxymethylated form. The N terminus of this protein was an N-acetylserine residue. The B-4 protein contained seven S-carboxymethylcysteine residues, six of which are located in the N-terminal region (residues 1-26), and other one in C terminus. The central region of the peptide chain was rich in hydrophobic residues. There were homologous amino acids at 66 positions in the sequences of the feather keratins of fowl, emu and silver gull. The variation (substitution, deletion and insertion) in sequence was found to be localized in both terminal sections of the polypeptide chain. The B-4 protein structure was predicted to contain beta-sheet (about 30%), turn and random-coil-like structure, and no alpha-helix. beta-Sheet structure is mostly located in the central region (residues 22-70). On the other hand, both terminal regions are almost devoid of secondary structure.

Amino Acid Sequence↗

Regularities and irregularities in the structure of the seminiferous epithelium in the domestic fowl (Gallus domesticus). I. Suggestion of the presence of the seminiferous epithelial cycle.

A cellular association demarcated by two perpendiculars which were drawn between adjacent bundles of elongate spermatids from the tubular lumen to the basement membrane, was made the unit of histometrical observation in this study (provisionally called a "column"). Cell counting revealed that the average numbers per column of various types of germ cells do not show any significant differences among 5 fowls and between paired testes. The frequency of spermiogenic steps (numbered 1-8) was investigated in each column. A definite and common pattern was found in the frequency distribution in the 5 fowls observed. A relationship between spermiation and younger spermatid steps was also investigated in each column. The spermiation was found at different steps, but most frequently at step 2 (30.6%). Based on these observations and referring to other author's information, an average time interval between two successive spermiations was calculated roughly at 3.3 +/- 1.2 days. Theoretically, this value is equal to an average length of one epithelial cycle. Such a variable cycle may have caused irregular cellular associations in this species.

Animals↗

A seminoma in a monorchid guinea fowl (Numida meleagris).

A case of seminoma in a monorchid adult guinea fowl (Numida meleagris) is described. Grossly, a right enlarged testis, which was soft in consistency, and white to pale in colour with few spots of haemorrhages was observed. Histologically, the testicle revealed diffusely spread sheets of tumour cells. The cells were large pleomorphic with eccentrically placed hyperchromic nuclei. Mitotic figures were evident. A scanty fibrous stroma, containing lymphocytes and histiocytes, separating the groups of tumour cells, along with few areas of haemorrhages were observed. Occurrence of seminoma in guinea fowl is unusual and hence reported.

Animals↗

Evidence that adrenaline is released from adrenergic neurones in the rectum of the fowl.

1 The rectum isolated from the fowl was perfused with Tyrode solution via the caudal mesenteric artery. Noradrenaline and adrenaline were biologically or fluorimetrically assayed in perfusates collected before and during stimulation of Remak's nerve or of the periarterial nerves.2 Perfusates collected during nerve stimulation relaxed the chick rectum and rat stomach strips which served as assay tissues. This effect was attributed to the action of noradrenaline or adrenaline released from adrenergic nerve endings which appeared in the perfusates.3 Perfusates obtained during stimulation (30 Hz for 60 s) of Remak's nerve contained both noradrenaline and adrenaline when measured fluorimetrically. The mean output per stimulus train was 0.8 +/- 0.2 ng/g wet wt. tissue for noradrenaline and 1.7 +/- 0.2 ng/g wet wt. tissue for adrenaline (n = 7). Perfusates obtained during stimulation (30 Hz for 60 s) of the periarterial nerves contained noradrenaline in a concentration of 1.6 +/- 0.3 ng/g wet wt. tissue per stimulus train, but almost no adrenaline (n = 7).4 Neither stimulation of Remak's nerve nor the periarterial nerves liberated catecholamines when the rectum was perfused with Tyrode solution containing low Ca(2+) (0.1 mM) and high Mg(2+) (10 mM).5 Infusion of high potassium solution (50 mM) increased markedly the output of noradrenaline and adrenaline.6 Adrenaline as well as noradrenaline may function as the adrenergic neurotransmitter in the rectum of the fowl.

Animals↗

Characterisation of Pasteurella multocida isolated from fowl cholera outbreaks on turkey farms.

Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate Pasteurella multocida isolates from outbreaks of fowl cholera on 7 turkey farms in New South Wales. While only a single isolate was available from 5 of the farms, multiple isolates, 4 and 12 respectively, were available from the other 2 farms. The available field evidence suggested that 8 outbreaks had occurred with one farm suffering 2 outbreaks. The isolates obtained were all confirmed as Pasteurella multocida. Biochemical profiles allocated the isolates to 4 groups, 3 being variants of P multocida subsp multocida and the fourth being P multocida subsp septica. REA performed with HpaII established 7 groups. Ribotyping using the HpaII digests probed with the 16S rRNA operon of Haemophilus paragallinarum recognised the same 7 groups as REA. Unlike the biochemical profiles, both REA and ribotyping provided a fine subdivision that identified outbreaks as either related or unrelated. The REA and ribotyping patterns as well as biochemical profiles were stable for all isolates from the outbreaks in which multiple isolates were obtained from either the same bird or from different birds. REA and ribotyping were found to be superior to biotyping methods for the investigation of fowl cholera outbreaks.

Animals↗

Water diuresis in the domestic fowl.

1. The exteriorization of ureters in domestic fowls allows the collection of urine uncontaminated by faeces.2. When water is administered by stomach tube to a fowl, water is stored in the crop, from which it is slowly released into the proventriculus.3. Immediately after water gavage, there is a sharp rise of urine flow accompanied by an increase in endogenous creatinine excretion and a fall in the osmolarity of the urine. This initial increase in urine flow is of short duration, and is followed by a normal curve of diuresis with a peak at about 90 min after hydration.4. The first rise in urine flow following the filling of the crop appears to be of reflex origin, as distension of the crop with paraffin produces an enhanced excretion of urine and of creatinine.5. Changes in urine flow and in creatinine excretion are closely correlated.6. After water deprivation for 18 hr, the urine is slightly hyptertonic to blood. The maximum urine concentration observed was of the order of 600 m-osmole/l.

Animals↗

Chromosomal translocation in domestic fowl induced by x-rays.

The cytological appearance and behavior of an x-ray-induced reciprocal translocation between the first and second chromsome of the domestic fowl is described, and its relevance to the further definition of linkage studies in the fowl is observed.

Animals↗

Fowl cholera immunization in turkeys. I. Efficacy of various cell fractions of Pasteurella multocida as vaccines.

Cell fractions of Pastuerella multocida (P-1059) were tested as vaccines against fowl cholera in turkeys. These fractions were culture filtrate, cell wall, and cytoplasm. A second culture filtrate preparation made from cells grown on blood-agar rather than the standard medium was also tested along with a "combination" preparation made by recombination of the cell fractions. Each preparation was tested in three vehicles: saline, alum (0.5%), and Freund Incomplete Adjuvant (50%). The turkeys vaccinated with these preparations were challenged by exposure to an experimental epornitic of fowl cholera. The combination fraction appeared to be the most promising vaccine when compared to the protective action of the commercial bacterin included in the test as a positive control.

Agar↗

Specific detection of Pasteurella multocida in chickens with fowl cholera and in pig lung tissues using fluorescent rRNA in situ hybridization.

A Pasteurella multocida species-specific oligonucleotide probe, pmhyb449, targeting 16S rRNA was designed and evaluated by whole-cell hybridization against 22 selected reference strains in animal tissues. It differentiated P. multocida from other bacterial species of the families Pasteurellaceae and Enterobacteriaceae and also from divergent species of the order Cytophagales (except biovar 2 strains of Pasteurella avium and Pasteurella canis, which have high 16S rRNA similarity to P. multocida). The potential of the probe for specific identification and differentiation of P. multocida was further detected in formalin-fixed paraffin-embedded lung tissues from experimental fowl cholera in chickens and infections in pigs. In chicken lung tissues P. multocida cells were detected singly, in pairs, as microcolonies, and as massive colonies within air capillaries (septa and lumen), parabronchial septa, and blood vessels (wall and lumen). In pig lung, postmortem-injected P. multocida was detected in the alveoli (lumen and wall), and in both animals the bacterial cells were seen in the bronchi. The results showed that with the oligonucleotide probe pmhyb449, fluorescent in situ hybridization is a suitable and fast method for specific detection of P. multocida in histological formalin-fixed tissues. The test was replicable and reproducible and is recommended as a supplementary test for diagnosis and as a tool in pathogenesis studies of fowl cholera and respiratory tract infections in pigs due to P. multocida.

Animals↗

Inhibition of cellular protein synthesis by simultaneous pretreatment of host cells with fowl plague virus and actinomycin D: a method for studying early protein synthesis of several RNA viruses.

A method is described for analysis of viral protein synthesis early after infection when minute amounts of viral proteins are effectively concealed by large amounts of produced host-specific proteins. The method is superior to a radioimmune assay, since all virus-induced proteins can be measured independent of their immunological reactivity. Host-specific protein synthesis can be suppressed by infection with fowl plague virus. Addition of actinomycin C 1.25 h postinfection does not prevent this suppression, but it does block effectively the formation of fowl plague virus-specific proteins. Such cells synthesize only small amounts of cellular proteins, as revealed by polyacrylamide electrophoresis. They can be superinfected with several different enveloped viruses, however, without significant diminution of virus yeilds. In pretreated cells the eclipse is shortened for Semliki Forest virus, Sindbis virus, and vesicular stomatitis virus, but prolonged for Newcastle disease virus. The onset of protein synthesis, specific for the superinfecting virus, could be clearly demonstrated within 1 h after superinfection. At this time, in cells superinfected with Semliki Forest virus, great amounts of NSP 75 (nonstructural protein; molecular weight, 75 X 10(3)) and reduced amounts of the core protein C could be deomonstrated. The precursor glycoprotein NSP 68 is followed by a new polypeptide, NSP 65: three proteins with molecular weights exceeding 100 X 10(3) were observed which are missing later in the infectious cycle. Similar results were obtained after superinfection with Sindbis virus. The formation of a new polypeptide with a molecular weight of about 80 X 10(3) was detected. After superinfection with vesicular stomatis virus or Newcastle disease virus the formation of new proteins, characteristic for the early stage of infeciton, was not observed.

Cell Line↗

Rescue of vector-expressed fowl plague virus hemagglutinin in biologically active form by acidotropic agents and coexpressed M2 protein.

The hemagglutinin of the Rostock strain of fowl plague virus was expressed in CV-1 cells by a simian virus 40 vector, and its stability in the exocytotic transport process was examined by a fusion assay. A 50-fold increase in the fusion activity of the hemagglutinin was observed when expression occurred in the presence of ammonium chloride, Tris-HCl, or high doses of amantadine. When chloroquine, another acidotropic agent, was used, the hemagglutinin exposed at the cell surface had to be activated by trypsin, because intracellular cleavage was inhibited by this compound. Hemagglutinin mutants resistant to intracellular cleavage did not require acidotropic agents for full expression of fusion activity, when treated with trypsin after arrival at the cell surface. These results indicate that fowl plague virus hemagglutinin expressed by a simian virus 40 vector is denatured in the acidic milieu of the exocytotic pathway and that cleavage is a major factor responsible for the pH instability. Coexpression with the M2 protein also markedly enhanced the fusion activity of the hemagglutinin, and this effect was inhibited by low doses of amantadine. These results support the concept that M2, known to have ion channel function, protects the hemagglutinin from denaturation by raising the pH in the exocytotic transport system. The data also stress the importance of acidotropic agents or coexpressed M2 for the structural and functional integrity of vector-expressed hemagglutinin.

Ammonium Chloride↗

A case report of fowl plague keratoconjunctivitis.

A case of human fowl plague keratoconjunctivitis occurred after accidental laboratory exposure. The conjunctivitis was characterised by follicle formation and a mucopurulent discharge, and ran a self-limiting course over two weeks. The keratitis was of an unusual type and consisted of small intraepithelial opacities, which appeared after one week and resolved completely over the next three weeks. The infection, confirmed by viral culture, was produced by Dutch strain (Hav 1 Neq 1) of fowl plague virus.

Adult↗