The role of retinal and extra-retinal photostimulation in reproductive activity in broiler breeder hens

Photostimulation of retinal photoreceptors, which are sensitive to green light, appears to inhibit reproductive activity in birds, whereas photostimulation of extra-retinal photoreceptors, which are sensitive to red light, accelerates it. The objective of this study was to determine the effect of either retinal or extra-retinal photostimulation on reproductive activities of broiler breeder hens. At 23 wk of age, Cobb hens (N = 135) were divided into 9 rooms with individual cages (n = 15). At 24 wk of age, 3 rooms were photostimulated (14L:10D) with white light (Control, n = 45). Six rooms had 2 parallel lighting systems, red (660 nm) and green (560 nm), which were both on during 6 out of 14 h of the light period. Then, in 3 of these rooms, the green light was turned off and hens were exposed to a total of 14 h of red light (Red, n = 45), and in the other 3, the red light was turned off and green lighting continued for a total of 14 h (Green, n = 45). The Green group had reduced egg production; reduced plasma concentrations of ovarian steroids; reduced luteinizing hormone (LH)-β, vasoactive intestinal peptide (VIP), and prolactin mRNA expression; and greater retinal green opsin mRNA expression (P ≤ 0.05). The Red group had greater egg production; greater gonadotropin-releasing hormone-I (GnRH-I) and red opsin gene expression in the hypothalamus; and lesser green opsin gene expression in the retina (P ≤ 0.05). We suggest that selective photostimulation of extra-retinal photostimulation as opposed to retinal photostimulation is a key factor in the determination of successful reproduction of broiler breeder hens.     

Photokinesis and Djopsin gene expression analysis during the regeneration of planarian eyes

Planarians provide the ideal model for studying eye development, with their simple eye structure and exceptionally rapid regeneration. Here, we observed the eye morphogenesis, photophobic behavior, spectral sensitivity and expression pattern of Djopsin in the freshwater planarian Dugesia japonica. The results showed that: (i) Djopsin encoding the putative protein belonged to the rhabdomeric opsins group and displayed high conservation during animal evolution; (ii) planarians displayed diverse photophobic response to different visible wavelengths and were more sensitive to light blue (495 nm) and yellow (635 nm); (iii) the morphogenesis and functional recovery of eyes were related to the expression pattern of Djopsin during head regeneration; and (iv) Djopsin gene plays a major role in functional recovery during eye regeneration and visual system maintenance in adult planarians.     

微量间接血凝试验检测特种经济动物嗜肺性军团菌抗体的研究

应用嗜肺性军团菌(Lp)共同抗原建立了微量间接血凝试验(MPHA),并以此方法对马鹿、梅花鹿、紫貂、水貂、貉、獭兔、熊、蓝狐、银狐及雉鸡等10种特种经济动物的830份血清样本进行了Lp抗体水平的检测。结果表明,紫貂和水貂血清的Lp抗体滴度1:32为阻抑试验阳性,而梅花鹿、马鹿、貉等8种特种经济动物血清的Lp抗体滴度1:64为阳性。按此标准,獭兔血清Lp抗体的阳性率为30.0％(9/30),梅花鹿15.3％(19/124),紫貂8.7％(2/23),马鹿8.45％(6/71),貉2.94％(6/204),水貂1.1％(8/273)。熊、蓝狐、银狐及雉鸡等血清样本全部呈阴性反应。     

Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells

The present study was carried out to examine the effects of post‐activation treatment of trichostatin A (TSA), a histone deacetylase inhibitor, on in vitro development and transgene function of somatic cell nuclear transfer (SCNT) embryos derived from Clawn miniature pig embryonic fibroblast (PEF) transfected with a bacterial endo‐β‐galactosidase C gene (removal of the α‐galactosyl (Gal) epitope). SCNT embryos were incubated with or without TSA (50 or 100 nmol/L) after activation, cultured in vitro and assessed for cleavage, blastocyst formation and transgene function. The rate of blastocyst formation was significantly higher in SCNT embryos treated with 50 nmol/L TSA than that in control (P < 0.05), whereas the rate of cleavage and cell number of blastocyst did not differ. Following labelling with fluorescein isothiocyanate‐labelled BS‐I‐B₄ isolectin, the intensity of fluorescence observed on cell‐surface was dramatically reduced in transgenic SCNT blastocyst in comparison with non‐transgenic SCNT blastocyst. However, the reduction of α‐Gal epitope expression in transgenic SCNT blastocyst was not affected by TSA treatment. The results of this study showed that post‐activation treatment with 50 nmol/L TSA is effective to improve in vitro developmental capacity of transgenic SCNT miniature pig embryos without the modification of transgene function.     

Tyrosine‐derived 4‐hydroxyphenylpyruvate reacts with ketone test fields of 3 commercially available urine dipsticks

Background: The enzyme 4‐hydroxyphenylpyruvate dioxygenase (HPPD) is key in tyrosine catabolism. Inhibition of HPPD results in tyrosinemia and increased urinary excretion of 3 phenylketones: 4‐hydroxyphenylpyruvate (HPPA), 4‐hydroxyphenyllactate (HPLA), and 4‐hydroxyphenylacetate (HPAA). A previous study involving administration of a novel HPPD inhibitor to dogs resulted in detection of ketonuria in treated animals using urine dipsticks read by reflectance photometry. Dipstick‐positive results were suspected to be false because high concentrations of urinary phenylketones have been reported to react with ketone test fields of urine dipsticks, but visual confirmation was not performed. Objective: The purpose of this study was to determine which of the 4‐hydroxyphenolic acids produced by HPPD inhibition react with ketone test fields of 3 commercially available urine dipsticks. Methods: Canine urine samples were prepared with HPPA, HPLA, HPAA, and lithium acetoacetate (positive control) at 6 concentrations. Unmodified urine samples were used as negative controls. All samples were tested for ketones using Combur 10 Test M dipsticks read by a Miditron dipstick analyzer. Urinalysis was also performed by visually inspecting ketone test fields on the Combur 10 Test M, Multistix 10 SG, and Aution 10 EA dipsticks. Results: Urine samples containing HPPA were positive for ketones with Combur 10 Test M dipsticks read by the Miditron analyzer and produced a red–brown color change in ketone test fields of all 3 dipsticks. Urine samples containing HPLA and HPAA were negative by all methods. Conclusion: The phenylketone HPPA reacts with ketone test fields of 3 commercially available urine dipsticks, producing a red–brown color change that may be misinterpreted as positive for ketones by reflectance photometry.     

Comparative morphology of the tapetum lucidum (among selected species)

Objectives The phenomenon of ‘eye‐shine’ is seen in a variety of animal species, and is generally thought to be related to the presence of an intraocular reflecting structure, the tapetum lucidum. The tapetum lucidum is a biologic reflector system that is a common feature in the eyes of vertebrates. It normally functions to provide the light‐sensitive retinal cells with a second opportunity for photon‐photoreceptor stimulation, thereby enhancing visual sensitivity at low light levels. The tapetum lucidum is presented here according to a classification based on the location, as well as the composition, of this reflective layer. Finally, the physical and chemical properties, as well as the origins of the different tapeta lucida, are discussed and compared. Methods The anatomic and biochemical aspects of the tapetum lucidum in various vertebrates are examined. Morphologic observations were made from paraffin and plastic embedded specimens. Specimens were treated with traditional stains and observed by light and transmission electron microscopy. Results Some species (primates, squirrels, birds, red kangaroo and pig) do not have this structure and they usually are diurnal animals. In vertebrates, the tapetum lucidum exhibits diverse structure, organization and composition. Therefore, the retinal tapetum (teleosts, crocodilians, marsupials, fruit bat), the choroidal guanine tapetum (elasmobranchs), the choroidal tapetum cellulosum (carnivores, rodents, cetacea), and the choroidal tapetum fibrosum (cow, sheep, goat, horse) are described. Conclusions The tapetum lucidum represents a remarkable example of neural cell and tissue specialization as an adaptation to a dim light environment and, despite these differences, all tapetal variants act to increase retinal sensitivity by reflecting light back through the photoreceptor layer. These variations regarding both its location and structure, as well as the choice of reflective material, may represent selective visual adaptations associated with their feeding behavior, in response to the use of specific wavelengths and amount of reflectance required.     

Phenotypic and molecular characterization of isolates of Ornithobacterium rhinotracheale from chickens and pigeons in Taiwan

Forty Ornithobacterium rhinotracheale (ORT) strains were isolated from 28 chickens and 12 pigeons for the first time in Taiwan. All isolates reacted positively in the p-nitrophenyl-beta-D-galactopyranoside (PNPG) and oxidase tests, showing an API 20NE identification system biocode 0-0-2-0-0-0-4. All the pigeon isolates and 85.7% (24 of 28) of the chicken isolates belonged to serotype A. Compared to the ORT ATCC 51464 strain, 14.3% (4 of 28) of chicken isolates and 58.3% (7 of 12) of pigeon isolates showed smaller colonies after 72 hr incubation. Most of the chicken isolates (22 of 28), but none of the pigeon isolates, could agglutinate chicken and pigeon red blood cells. There appears to be a correlation that ORT isolates with a larger colony size tend to be more able to agglutinate red blood cells than the ORT isolates with a smaller colony size. A majority of isolates was sensitive to amoxicillin, ampicillin, ceftiofur, penicillin, and oxytetracycline. The 16S ribosomal RNA (rRNA) sequences of 23 Taiwanese ORT isolates showed high identity (98%-100%) to sequences in GenBank. Phylogenetic analysis of these sequences showed that pigeon isolates formed a distinctive cluster, while chicken isolates and all other 16S rRNA sequences obtained from GenBank belonged to another two clusters. The results indicate that pigeon ORT isolates are different from most chicken isolates in regard to a number of phenotypic and molecular traits.     

Red‐light stimulation of boar semen prior to artificial insemination improves field fertility in farms: A worldwide survey

A survey of in vivo fertility data from 31 pig farms distributed worldwide was conducted to determine whether stimulating boar semen with LED‐based red light increases its reproductive performance following artificial insemination (AI). Red‐light stimulation with MaXipig^® was found to increase farrowing rates (mean ± SEM, control: 87.2% ± 0.4% vs. light stimulation 90.3% ± 0.5%) and the number of both total and live newborn piglets. Red‐light stimulation increased farrowing rates in 27 farms, with an increase ranging from 0.2% to 9.1%. Similar results were observed in litter sizes. Suboptimal management after AI was suggested in those farms with no response to red‐light stimulation. Our results indicate that a routine use of red‐light stimulation of boar semen can have a positive effect on the reproductive performance. However, the effectiveness of this system appears to highly rely upon proper management of pig farms.     

黑化雉鸡羽色遗传的研究

通过黑化雉鸡 (Melanisticmuckedpheasant)和大型环颈雉鸡 (Jumboringneckedpheas ant)的正交和反交 ,以及F1 中进行自交 ,观察F1 和F2 中羽色性状的表现和分离比例。试验结果表明 ,黑化雉鸡羽色性状对于环颈雉鸡羽色的遗传方式是显性遗传 ,而且显性是完全的 ;黑化雉鸡羽色性状也由常染色体上一对等位基因M来控制的 ,并不存在显性致死现象     

Leptospiral lipopolysaccharide stimulates the expression of toll‐like receptor 2 and cytokines in pig fibroblasts

Pigs throughout the world are afflicted with leptospirosis, causing serious economic losses and potential hazards to human health. Although it has been known that leptospiral lipopolysaccharide (L‐LPS) is involved in an immunological reaction between an antigen and a host cell, little is known about how the immune system of pigs can respond to L‐LPS. Here, we stimulated pig fibroblasts by L‐LPS and then quantitatively measured gene and protein expression levels of two toll‐like receptors (TLRs), TLR2 and TLR4, by real‐time PCR and Western blotting. As a result, expression of TLR2 was found to be significantly up‐regulated within 24 h after L‐LPS stimulation whereas induction of TLR4 expression was relatively weak. We also revealed that of myeloid differentiation primary response gene 88 (MyD88), interleukin 6 (IL‐6) and IL‐8 gene expressions were markedly up‐regulated by L‐LPS stimulation. These results may suggest that the pig cell can activate TLR2 rather than TLR4 by L‐LPS stimulation, thereby inducing expression of cytokines.     

The presence of megamitochondria in the ellipsoid of photoreceptor inner segment of the zebrafish retina

Although the megamitochondria (MM) were localized in various pathological conditions, normal retina of some mammalian species was reported to include MM for various physiological roles. However, it was not clearly confirmed whether the MM is present in the retina of lower vertebrate as well. In this study, we tried to show the presence of the MM in the zebrafish retina using electron microscopic technique. In all the photoreceptors including rods, cones and double cones of the zebrafish retina, MM were observed in the ellipsoid of inner segment. In the photoreceptor epllipsoid of the zebrafish retina, the mitochondria located in the central portion of the ellipsoid had a highly electron-dense matrix, which were accompanied by the mitochondria with electron-lucent matrix in the apical portion of the ellipsoid. The presence of MM was more clearly discernable in the rods, which were localized under the double cones. This finding is somewhat different from those observed in the previous studies because MM were localized in the inner segment of cones, but were not in those of rods in the case of mammalian retina. Although the exact physiological meaning for the presence of MM in some vertebrate species should be further studied, the present study could show that the MM in the ellipsoid of the retinal photoreceptors was not only restricted in some mammalian species.     

Isolation and Identification of Bacterial Pathogen of Pig Arthritis in Yunnan

This experiment was conducted to investigate the bacterial causes and drug resistance of pig arthritis of four farms in Yunnan.Through isolation and culture,biochemical test,drug sensitive test,animal pathogenicity experiment and 16S rRNA identification,we analyzed the samples from four farms with pig arthritis symptom.The results showed that in all 96 samples,55 samples were Staphylococcus aureus positive,the positive rates were 20.8%,83.3%,91.7% and 33.3% in A,B,C and D farms,respectively;56 samples were Streptococci positive,the positive rates were 95.8%,21.7%,25.0% and 83.3%,respectively;Mixed infection rates were 20.8%,21.7%,25.0% and 33.3%,respectively.Therefore,Staphylococcus aureus was the major bacterial pathogen which caused pig arthritis in B and C farms;Streptococci was the major bacterial pathogen which caused pig arthritis in A and D farms;Two kinds of bacteria were more sensitive to ceftriaxone,minocycline and ciprofloxacin,and resistant to multiple antibiotics.     

云南猪关节炎细菌性病原的分离鉴定

本试验旨在了解云南4个猪场猪关节炎的细菌性病因及其耐药性.通过分离培养、生化试验、药敏试验、动物致病性试验及16S rRNA鉴定,对4个猪场出现猪关节炎症状的样品进行了分析.结果显示,A、B、C和D 4个猪场共96份样品中,检出金黄色葡萄球菌55份,阳性率分别为20.8%、83.3%、91.7%和33.3%;检出链球菌56份,阳性率分别为95.8%、21.7%、25.0%和83.3%;混合感染阳性率分别为20.8%、21.7%、25.0%和33.3%.因此,金黄色葡萄球菌是导致B、C两猪场猪关节炎的主要细菌性病原;链球菌是导致A、D两猪场猪关节炎的主要细菌性病原;两种细菌对菌必治、米诺环素和环丙沙星较敏感,对多种抗生素产生了耐药性.     

Epidemiology of Chlamydia psittaci Infection in Racing Pigeons and Pigeon Fanciers in Beijing,China

Over 3 million racing pigeons (Columba livia) are registered in Beijing City Center for gambling purposes. During 2008–2010, we evaluated the occurrence and prevalence of Chlamydia psittaci in racing pigeons as well as the possible zoonotic transmission to pigeon fanciers in six districts of Beijing where pigeon races are particularly popular. C. psittaci‐specific serum antibody titres were obtained from 370 pigeons and 79 fanciers using enzyme‐linked immunosorbent assay. In addition, 206 and 67 throat swabs were, respectively, collected from pigeons and fanciers and tested for the presence of chlamydial antigen using immunofluorescence. C. psittaci‐specific serum antibody was detected in 37 of 370 pigeons and 19 of 79 fanciers. Of 206 pigeon clinical specimens, 55 were positive for C. psittaci antigen, while 16 of 67 swabs from the pigeon fanciers were positive. Based on ompA sequence analysis, the genotype of several avian and human isolates was genotype B. Thus, both high‐titre C. psittaci‐specific antibody and C. psittaci‐specific antigen were found with relatively high frequency in the pigeon flocks as well as in the pigeon fanciers. Our study suggests that C. psittaci infection is prevalent among the racing pigeon population in Beijing. Moreover, detection of serum antibodies and antigen in pigeon fanciers suggests that exposure and possible zoonotic transmission of C. psittaci from racing pigeons to humans does occur. In view of the life‐threatening respiratory illness C. psittaci may cause in humans, regulatory public health measures, to prevent further spread of the pathogen in avian populations and possible transmission to exposed humans, are urgently needed.     

Characteristics of structures and lesions of the eye in laboratory animals used in toxicity studies

Histopathology of the eye is an essential part of ocular toxicity evaluation. There are structural variations of the eye among several laboratory animals commonly used in toxicity studies, and many cases of ocular lesions in these animals are related to anatomical and physiological characteristics of the eye. Since albino rats have no melanin in the eye, findings of the fundus can be observed clearly by ophthalmoscopy. Retinal atrophy is observed as a hyper-reflective lesion in the fundus and is usually observed as degeneration of the retina in histopathology. Albino rats are sensitive to light, and light-induced retinal degeneration is commonly observed because there is no melanin in the eye. Therefore, it is important to differentiate the causes of retinal degeneration because the lesion occurs spontaneously and is induced by several drugs or by lighting. In dogs, the tapetum lucidum, a multilayered reflective tissue of the choroid, is one of unique structures of the eye. Since tapetal cells contain reflecting crystals in which a high level of zinc has been demonstrated chemically, drug-induced tapetum degeneration is possibly related to zinc chelation. The eye of the monkey has a macula similar to that of humans. The macula consists only of cones with a high density, and light falls directly on the macula that plays an important role in visual acuity. Macular degeneration occurring in monkeys resembles histopathologically that of humans. Hence, the eye of the monkey is a suitable model to investigate macular degeneration and to assess drug-induced macular lesions.     

Isolation,Identification and Analysis of Serotype and Drug Resistance of Salmonella from Pigeon in Shanghai

The purpose of this study was to investigate the molecular types and antimicrobial resistance of Salmonella isolated from pigeon in Shanghai from 2011 to 2014. A total of 92 fecal samples were collected from markets. XLD plate and Salmonella chromogenic medium were used to isolate suspected Salmonella colonies, and then determined the number of Salmonella through gram staining and biochemical tests. Among them, 24(26.1%) were positive for Salmonella. Kauffmann-White method and the K-B method were used respectively for serotype identification and susceptibility testing. In result, serological identification showed that 24 isolates from pigeon could be divided into 3 serotypes including S.typhimurium (66.7%), S.agona (25.0%) and S.corvallis (8.3%). Drug susceptibility test showed that 75.0% of the isolates were resistant to one antibiotic at least. The highest level resistance were found for tetracycline as well as sulfisoxazole (62.5%), followed by streptomycin (58.3%), nalidixic acid (50.0%) and ampicillin (20.8%). Multi-drug resistant strains accounted for 62.5% (15), in which the largest number of strains (7, 16.7%) were resistant to four drugs. In addition, isolates were 100.0% susceptible to cefepime, cefotaxime, ceftazidime, imipenem and ofloxacin, but 12.5% were moderately sensitive to the amoxicillin/clavulanic acid and ciprofloxacin. The study showed that Salmonella had a high separation rate of pigeon in Shanghai farmers market and performed serious multidrug resistance, which would bring great challenges and risks to the prevention and control of Salmonella in food, so that the prevalence and drug resistance of Salmonella in pigeon deserved close attention.     

上海市肉鸽沙门氏菌的分离鉴定及流行血清型和耐药性分析

为了解上海市农贸市场肉鸽群体中沙门氏菌流行血清型和耐药情况,本试验于2011-2014年间从上海市各区农贸市场采集肉鸽新鲜粪便样本92份,用XLD平板和沙门氏菌属显色培养基分离疑似沙门氏菌,革兰氏染色镜检并进行生化试验,共获得沙门氏菌24株,分离率为26.1%。采用Kauffmann-White法和K-B纸片法对24株沙门氏菌进行血清型鉴定和16种抗生素的药敏试验。血清型鉴定显示,24株沙门氏菌可分为鼠伤寒（66.7%）、阿贡纳（25.0%）和科瓦利斯（8.3%）3种血清型。药敏试验结果显示,有75.0%（18株）的分离株表现出不同程度耐药,其中分离株对四环素和磺胺异唑耐药率最高,均达到62.5%,其次为链霉素（58.3%）、萘啶酸（50.0%）、氨苄西林（20.8%）。多重耐药菌株15株（62.5%）,耐4种抗菌药物的菌株最多,占16.7%（7株）。另外,分离菌株对头孢吡肟、头孢噻肟、头孢噻甲羧肟、亚胺培南、氧氟沙星等5种药物的敏感率为100.0%,对奥格门丁和环丙沙星有12.5%的中介敏感率。本研究结果表明,上海市肉鸽中沙门氏菌的携带率较高,且菌株多重耐药现象较严重,这为食品中沙门氏菌的防控工作带来较大的挑战和风险,肉鸽沙门氏菌的流行和耐药情况应值得密切关注。     

应用平板凝集试验诊断雉鸡结核病的研究

本项研究应用平板凝集试验诊断雉鸡结核病，用禽型结核快速平板凝集反应抗原，检测雉鸡血清中的抗结核杆菌抗体，结果表明，本试验方法特异性强，敏感性高，检出率高，速度快，简便易行，适于现场大批检疫，从而解决了雉鸡结核活体不能诊断的难题，为防治雉鸡结核病提供了可靠的血清学诊断方法。     

Immunohistochemical studies of cone photoreceptors and cells of the inner retina in feline rod-cone degeneration

Previous studies using electroretinography and immunohistochemistry have shown normal cone function and structure in early stages of hereditary rod‐cone degeneration of Abyssinian cats. To further investigate the cone photoreceptors and the inner retina of dystrophic cats, antibodies against green‐ and blue‐sensitive cones and specific cell types of inner retina were used in seven cats with the recessively inherited rod‐cone degeneration, and three normal European short‐haired cats. There was a reduction in number of both types of cones early in the disease. Changes at early stages of disease also occurred among horizontal cells in which there was an extension and a thickening of their lateral processes. The regular configuration of bipolar cells was changed in the more advanced stages of disease and their apical dendrites were lost. Abnormalities were not observed in the amacrine cells and in the ganglion cell layer in any of the present cases. This study shows that the cone system is morphologically abnormal in young cats at an earlier stage of disease than previously shown. The present findings also support the assumption that the inner retina is largely preserved throughout the disease process.     

Adult-onset cerebellar cortical abiotrophy and retinal degeneration in a domestic shorthair cat.

A 4-year-old, neutered male domestic shorthair cat presented for evaluation of ataxia and visual deficits. Neurological examination revealed severe cerebellar ataxia with symmetrical hypermetria and spasticity, a coarse whole-body tremor, positional vertical nystagmus, and frequent loss of balance. A menace response was absent bilaterally, and the pupils were widely dilated in room light. A funduscopic examination revealed markedly attenuated to absent retinal vessels and pronounced tapetal hyperreflectivity, findings consistent with end-stage retinal degeneration. Blood work evaluation included retroviral testing, a complete blood count, serum biochemistry analysis, taurine levels, and toxoplasma immunoglobulin G and immunoglobulin M titers. All were within reference ranges. The patient was euthanized, and a necropsy was performed. Microscopically, lesions of the nervous system were confined to the cerebellum and were consistent with cerebellar cortical abiotrophy. Selective photoreceptor degeneration was seen on histopathological examination of the retina with a reduction in the number of rods and cones. The combination of clinical findings and histopathological lesions seen here has not been previously reported in the cat.