A survey of gastrointestinal helminths in cats of the metropolitan region of Rio de Janeiro, Brazil

The prevalence of gastrointestinal helminth parasites in 135 cats over 1 year of age and inhabiting the metropolitan region of the city of Rio de Janeiro, Brazil was investigated by necropsy. These animals had two distinct origins: 99 cats (29 males and 70 females) were derived by capture in public areas (feral/stray) and 36 (12 males and 24 females) from shelters. The overall prevalence of gastrointestinal helminth parasites was 89.6%. The following parasites, with their respective prevalence in parenthesis, were found: Dipylidium caninum (52.6%), Ancylostoma braziliense (65.9%), Ancylostoma tubaeforme (8.9%), Toxocara cati (25.2%), Toxascaris leonina (11.9%), Physaloptera praeputialis (9.6%). Concurrent infections with two or more parasites were recorded in 59.5% of the individuals. Of the 121 parasitized cats, 94 were feral/strays and 27 were from shelters. Among feral/stray cats, 80 were infected with A. braziliense (85%) and 17 of the shelter felids were infected with D. caninum (63%). Feral/stray cats had higher worm intensities (6411/94-68.2) than shelter cats (992/27-36.7). The helminth parasites most frequently found in feral/stray cats were Ancylostoma braziliense (47.5%) and D. caninum (47%) while in shelter cats, D. caninum was the predominant species (85.2%). Twenty seven cats (22.3%) had only A. braziliense and 19 (15.7%) were parasitized only with D. caninum. Among those cats harboring mixed infections A. braziliense and D. caninum were the species more frequently found (P < 0.001).     

Application of a species-specific PCR-RFLP to identify Ancylostoma eggs directly from canine faeces

Apart from their veterinary importance, the hookworms Ancylostoma caninum, Ancylostoma braziliense and Ancylostoma caninum are also capable of causing zoonotic disease in humans. A highly sensitive and species-specific PCR-RFLP technique was utilised to detect and differentiate the various canine Ancylostoma spp directly from eggs in faeces. This technique was utilised to screen 101 canine faecal samples from parasite endemic tea growing communities in Assam, India, as part as an ongoing epidemiological investigation into canine parasitic zoonoses. The prevalence of hookworms in dogs was found to be 98% using a combination of PCR and conventional microscopy. Overall, 36% of dogs were found positive for single hookworm infections with A. caninum, 24% positive for single infections with A. braziliense and 38% had mixed infections with both A. caninum and A. braziliense. No dogs were found positive for A. ceylanicum in the community under study. The high prevalence of A. caninum and A. braziliense in dogs in this community may account for the high incidence of cutaneous larva migrans (CLM) observed among the human population residing at the tea estates. The PCR-RFLP technique described herein allows epidemiological screening of canine hookworms to be conducted rapidly, with ease and accuracy, and has the potential to be applied to a number of different clinical, pharmacological and epidemiological situations.     

Efficacy of a chewable formulation of ivermectin against a mixed infection of Ancylostoma braziliense and Ancylostoma tubaeforme in cats.

The efficacy of a beef-based, chewable formulation of ivermectin against a mixed infection of Ancylostoma braziliense and A tubaeforme was determined in cats. Ivermectin administered orally at approximately 24 micrograms/kg of body weight was 92.8% effective against adult A braziliense and 90.7% effective against adult A tubaeforme. The number of eggs per gram of feces had decreased 98.1% by 7 days after treatment. Clinical signs of hookworm disease also decreased after treatment. Location of adult parasites within the small intestine, percentage of infecting larvae that developed to the adult stage, and egg size in cats with infections of A braziliense and A tubaeforme were similar to those reported for cats with separate infections of either species.     

Hookworms of feral cats in Florida

Thirty feral cats (Felis catus) from Alachua county (northern Florida) and 30 from Palm Beach county (southern Florida) were examined for hookworms. Two species, Ancylostoma tubaeforme and Ancylostoma braziliense, were identified. Forty-five cats (75%) were infected with A. tubaeforme, with a mean intensity of 48 hookworms per cat. Twenty cats (33%) were infected with A. braziliense, with a mean intensity of 28 worms per cat. The prevalence of A. tubaeforme was greater than that of A. braziliense in Alachua (P=0.002) and Palm Beach (P=0.004) counties. The intensity of A. tubaeforme infections was higher in Palm Beach county than Alachua county (P=0.013). The intensities of A. tubaeforme and A. braziliense were positively correlated (increased together) in Palm Beach county (P=0.011). These hookworms have also been identified in bobcats (Felis rufus), gray foxes (Urocyon cinereoargenteus), and red foxes (Vulpes vulpes) in Florida. The prevalence of A. tubaeforme was significantly greater in feral cats than those reported in bobcats (P<0.001). The prevalence of A. braziliense was significantly greater in feral cats than in those reported in gray foxes (P=0.008). The hookworm that infects Florida panthers and bobcats, A. pluridentatum, was not found.     

Helminths in dogs belonging to people in a resource-limited urban community in Gauteng, South Africa

One hundred and sixty-four faecal samples, 148 adhesive tape swabs and 132 blood samples were collected from dogs in Boksburg, Gauteng, to assess the prevalence of helminth parasites in the area. Sixty-nine of these dogs were also necropsied and helminths recovered from the gastro-intestinal tracts. Ancylostoma caninum was the most common helminth and was present in 88% of the dogs, followed by Dipylidium caninum in 39% of dogs, Toxocara canis in 36%, Ancylostoma braziliense in 20%, Spirocerca lupi in 14%, Toxascaris leonina in 9%, Trichuris vulpis in 6%, Joyeuxiella pascualei in 6% and Taenia spp. in 4%. Microfilariae of Dipetalonema reconditum were found in 2% of the blood samples. The results of this study extend the geographic distribution of T. vulpis. With the exception of Spirocerca and Dipetalonema, all these helminths are potentially zoonotic, and may pose a threat to community health.     

Gastro-intestinal helminths of domestic dogs in the Republic of South Africa

Of the gastro-intestinal helminths found in 1 063 dogs from different parts of the country, Dipylidium caninum, the most prevalent cestode, occurred in 19,7% of the animals. Echinococcus granulosus was present in 10 dogs, 8 of which were from urban areas, and Taenia hydatigena in 69 dogs. Taemia multiceps was recovered from dogs only in the Cape Province and the Orange Free State. Toxocara canis was present in 81 of the necropsies carried out on 253 dogs from the Pretoria municipal area, while Ancylostoma spp. were present in 175 dogs. In the Pretoria area Ancylostoma caninum (11,9%) was more prevalent than Ancylostoma braziliense (6,2%).     

Helminths in dogs from a peri-urban resource-limited community in Free State Province,South Africa

This study focused on the helminth parasites of dogs in the Bloemfontein area of the Free State Province of South Africa. Samples were collected from 63 stray dogs destined for euthanasia. Complete sets of samples, consisting of blood, faeces, adhesive tape swabs and organs (heart, lungs and complete gastro-intestinal tract) from each dog were examined for helminth parasites. Dipylidium caninum was the most common helminth, and was recovered from 44% of dogs, followed by Taenia spp. (33%), Toxascaris leonina (32%), Ancylostoma caninum (27%), Toxocara canis (21%), Ancylostoma braziliense (19%), Spirocerca lupi (13%) and Joyeuxiella sp. (5%). Although human infection with S. lupi and Joyeuxiella sp. are unlikely, all the other helminths reported in this study are zoonotic, and therefore pose a threat to community health in Bloemfontein.     

Faecal helminth egg and oocyst counts of a small population of African lions (Panthera leo) in the southwestern Kalahari, Namibia

An endoparasite survey of a small pride of African lions (Panthera leo) was conducted at Intu Afrika Kalahari Game Reserve, southwestern Namibia, during winter and summer of 2003 and 2004, respectively. Overall, 23 fresh lion scats were collected opportunistically during fieldwork trials. A flotation technique was employed for the diagnosis of parasites. Three nematodes, Ancylostoma braziliense, Gnathostoma spinigerum and Uncinaria stenocephala and two coccidians, Toxoplasma gondii and Isospora felis were recorded. By using the McMaster method for quantification, a maximum number of 14866 oocysts per gram of faeces was obtained for I. felis during winter 2003. Endoparasite taxa carried by the different individuals in the pride were found to be related to their levels of association. Rates of infection were relatively low as a result of the habitat, semi-captive conditions and earlier sporadic deworming.     

Comparison of febantel tablets and Vercom paste against gastrointestinal nematodes of dogs.

Thirty adult dogs with naturally acquired gastrointestinal nematode infections were assigned at random to ten replicates and treated daily for 3 days with either a combination febantel/praziquantel (Vercom) paste, febantel tablets or placebo tablets. Numbers of hookworm and whipworm eggs after treatment were reduced similarly for both drug formulations when compared with pretreatment egg counts, whereas these counts increased in the controls. Vercom paste reduced the hookworm egg count by 99.9% and the whipworm egg count by 99.6%. The febantel tablet decreased the hookworm egg count by 99.9% and the whipworm egg count by 100%. As determined at necropsy, the controlled test efficacy against adult hookworms and whipworms was similar for the Vercom paste and the febantel tablets. The controlled test efficacies of Vercom paste against Ancylostoma caninum, Ancylostoma braziliense, and Trichuris vulpis were, respectively, 99.7%, 100% and 95.8% and those of febantel tablets were 98.2%, 100% and 99.7%. These results indicate that the nematocidal efficacy of febantel against these nematodes remains unchanged in these two formulations. No adverse reactions to either febantel tablets or to Vercom paste were observed.     

Immunohistochemical expression of MDR1‐Pgp 170 in canine cutaneous and oral melanomas: pattern of expression and association with tumour location and phenotype

Canine melanoma (CMM) more commonly affects the oral mucosa and the cutis. CMM shares several features with human melanomas (HMM), included resistance to a broad variety of antineoplastic chemotherapy agents. P‐glycoprotein 1 (Pgp) expression is a well‐recognized feature of multi‐drug resistance and the purpose of this study was to investigate its expression in treatment naïve CMM. We also investigated Pgp association with tumour location and histological features. Histology records of CMM were retrieved, including patients from 2012–2014. Twenty‐five cases of CMM were included in this study. Results revealed that Pgp is expressed in CMM and oral tumours were more likely to have a membranous Pgp expression (100%) than cutaneous tumours (66.6%) (P = 0.010). Cytoplasmic and nuclear Pgp expression could also be identified. Results of this study bring useful data that help in understanding one of the possible mechanisms responsible of intrinsic chemotherapy resistance in canine CMM.     

Activity of ivermectin against canine intestinal helminths

The anthelmintic activity of ivermectin was tested in 98 dogs against adult ascarids (Toxocara canis, Toxascaris leonina), hookworms (Ancylostoma caninum, A braziliense), and whipworms (Trichuris vulpis), and against experimentally induced infections (4th-stage larvae) of T canis and A caninum. Dosage levels tested were single subcutaneous injections of 50, 100, 200, or 400 micrograms/kg of body weight with appropriate vehicle-treated controls. A minimum of 4 (usually 5) dogs were tested with each parasite and dosage level. The lowest dosage level, 50 micrograms/kg, and all higher dosage levels expelled greater than 99% of the adult forms of both species of hookworms and intestinal larval forms of A caninum, as determined by worm counts at necropsy. A dosage level of 100 micrograms/kg was required to expel greater than 99% of whipworms and 200 micrograms/kg was necessary to expel adult (91%) and larval (97%) stages of T canis. Ivermectin was only marginally effective (34.2%, 46.2%, 69.2%, and 53.8%) against Toxascaris leonina at 50, 100, 200, and 400 micrograms/kg, respectively, and had no effect against occasional infections with the tapeworms, Dipylidium caninum (14 dogs) and Taenia spp (3 dogs).     

影响山羊产羔数lncRNA的筛选及功能分析

旨在分析高、低繁殖力川中黑山羊卵巢组织中长链非编码RNA（long non-coding RNA,lncRNA）的表达谱,探讨lncRNA与山羊生殖调控的关系,为解释lncRNA在山羊繁殖调控中的分子机制提供理论依据。本研究选择3胎以上的10头健康川中母羊（3.5~4.5岁）,将其分为高繁组和低繁组。高繁组为每胎产羔数在2头以上的母羊（n=6）,低繁组为每胎产羔数为1头的母羊（n=4）。同期发情后,采集高繁和低繁山羊的卵巢样品,并从中提取总RNA构建测序文库,通过HigSeq X ten平台进行测序,筛选出差异lncRNAs,通过对差异显著的lncRNAs与mRNAs位置关系以及结合能的判定预测顺式作用的靶基因,并对其进行GO和KEGG通路分析。最后,随机抽取6个差异lncRNAs进行实时荧光定量验证。结果表明,本研究共鉴定出168个差异表达lncRNAs,其中,163个lncRNAs在高繁组卵巢显著上调,5个lncRNAs在低繁组卵巢显著上调。筛选出的差异表达lncRNAs可能为调控山羊繁殖力的候选lncRNAs,包括ENSCHIG00000001479、ENSCHIG00000002617和ENSCHIG00000002622等。这些候选lncRNAs的靶基因（MYC、CDH2、FGF9、PPARGC1A等）主要富集于Jak-STAT、细胞黏附分子及AMPK等信号通路,ENSCHIG00000001479的靶基因MYC参与了Jak-STAT、TGF-β和MAPK通路;ENSCHIG00000002617和ENSCHIG00000002622的靶基因CDH2参与了细胞黏附分子通路。通过qRT-PCR验证发现,定量结果与测序结果基本一致。研究结果推断,ENSCHIG00000001479、ENSCHIG00000002617和ENSCHIG00000002622等lncRNAs可能对山羊生殖发育过程中的卵巢发育及排卵等进程具有重要的调控作用,认为鉴定出的差异表达lncRNAs与山羊产羔数有关。本研究利用RNA-Seq技术筛选高、低繁殖力山羊的差异表达lncRNAs,并对其进行GO和KEGG功能分析,为探讨山羊产羔数的繁殖机制提供更完善的转录组数据。     

Haemoglobin in normal and neoplastic canine mammary glands

Four types of globins for oxygen transport are known in vertebrates, and the haemoglobin is responsible for carrying oxygen in blood. In this study, we found that haemoglobin was also expressed in canine mammary glands. Samples were taken from 26 malignant mammary tumors, 16 normal mammary glands and 10 other normal tissues. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and mass spectrometry were used to investigate haemoglobin in mammary tissues. The results indicated that normal canine mammary glands expressed high levels of haemoglobin protein as shown by Coomassie blue staining. The identity of haemoglobin was confirmed by immunoblotting and mass spectrometry, and the mass spectrometry data revealed that both alpha-haemoglobin and beta-haemoglobin were expressed. Relative to normal mammary glands, the levels of haemoglobin expression in mammary tumors were lower. Our results also indicated that the haemoglobin was endogenously produced in mammary gland tissues and was not derived from the erythroid cells.     

Identification and Analysis of CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 in Cynomolgus Macaques

Cytochromes P450 (P450) are important for not only drug metabolism and toxicity, but also biosynthesis and metabolism of cholesterol and bile acids, and steroid synthesis. In cynomolgus macaques, widely used in biomedical research, we have characterized P450 cDNAs, which were isolated as expressed sequence tags of cynomolgus macaque liver. In this study, cynomolgus CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 cDNAs were characterized by sequence analysis, phylogenetic analysis and tissue expression pattern. By sequence analysis, these five cynomolgus P450s had high sequence identities (94–99%) to the human orthologs in amino acids. By phylogenetic analysis, each cynomolgus P450 was more closely related to the human ortholog as compared with the dog or rat ortholog. By quantitative polymerase chain reaction, among the 10 tissue types, CYP7A1 and CYP17A1 mRNAs were preferentially expressed in liver and adrenal gland, respectively. Cynomolgus CYP27A1 and CYP51A1 mRNAs were most abundantly expressed in liver and testis, respectively. Cynomolgus CYP20A1 mRNA was expressed in all the tissues, including brain and liver. Tissue expression patterns of each cynomolgus P450 were generally similar to that of the human ortholog. These results suggest the molecular similarities of CYP7A1, CYP17A1, CYP20A1, CYP27A1 and CYP51A1 between cynomolgus macaques and humans.     

初情期母牛下丘脑-垂体-卵巢转录组分析

通过对3头18月龄安格斯牛下丘脑-垂体-卵巢组织进行转录组测序和生物信息学比较分析,筛选和挖掘与肉牛繁殖活动相关候选基因和信号通路。通过转录组测序,进行表达基因分析、GO和KEGG富集分析以及信号通路筛选。结果显示,共获得61.92 Gb Clean Data,各样品Clean Data均达到6.88 Gb,Q30碱基百分比在92.42%及以上。有15154个基因在所有组织中都有表达,有4780个基因只在2种组织器官中共同表达,有5809个基因只在其中的一个组织器官中表达。下丘脑-垂体-卵巢各组织所表达的基因中,在GO数据库得到注释的分别为2168,2101,13777个,在KEGG数据库中得到注释分别有12477,12446,12176个。在所有KEGG通路中,Notch信号通路(notch signaling pathway)、神经营养因子信号通路(neurotrophin signaling pathway)和血管内皮生长因子信号通路(VEGF signaling pathway)的表达频率最高,说明这些信号通路可能也参与下丘脑-垂体-卵巢轴对母牛繁殖活动的调节。     

Expressed sequence tags from life cycle stages of Trichinella spiralis: application to biology and parasite control

While the approach taken to date to study Trichinella spp., involves mainly characterization of individual genes of interest, we initiated a genomics approach as an antecedent to more complete genome sequencing. Our approach involves use of expressed sequence tags (ESTs) obtained from three life cycle stages of Trichinella spiralis; adult worms (AD), mature muscle larvae (ML) and immature L1 larvae (immL1, also known as newborn larvae) () to improve the technical capacity for research on Trichinella spp. and to generate information that will aid prospective development of relevant hypotheses. In this review, we will summarize findings of our EST analysis and discuss how they relate to topics mentioned above. The foundation laid by this data will also contribute toward development of a more substantial genomic database and technical capacity to dissect molecular interactions between vertebrate hosts and Trichinella spp.     

2例犬肝胆管细胞癌的病理学诊断

肝胆管细胞癌是来源于肝胆管上皮细胞的恶性肿瘤,在多物种中都有发生。笔者运用组织病理学和免疫组织化学的方法,对2例犬肝肿瘤进行了诊断。结果显示：肝肿瘤眼观均为圆球形白色肿块,组织病理学检查肿瘤组织由呈腺管状排列的瘤细胞构成,有的腺管中央有坏死脱落的细胞,有的有黏液样物质,腺管间以薄的纤维结缔组织分隔,瘤组织有大片的坏死。瘤细胞多呈卵圆形,细胞核大、卵圆形,核分裂象多见,瘤细胞的CK19阳性表达,而CK18和AFP呈阴性表达。根据组织病理学和免疫组织化学检查结果判断,2例犬肝肿瘤为肝胆管细胞癌,本研究为犬肝肿瘤诊断积累了病理学资料。     

体细胞核移植技术生产猪植酸酶转基因胚胎的研究

为获得具有植酸酶腮腺特异性表达的猪转基因克隆胚胎,本研究使用植酸酶腮腺特异性表达的DNA质粒(包含腮腺分泌蛋白(parotid secretary protein,PSP)启动子与终止子序列、Neo筛选基因、绿色荧光蛋白(EGFP)报告基因和高比活的植酸酶appA基因),采用脂质体转染和基因素418(G418)药物抗性筛选的方法获取稳转细胞系,并利用体细胞核移植技术获得植酸酶转基因胚胎。结果表明,本研究构建的DNA质粒可用于细胞筛选,且质粒越小,细胞的转染效率越高,14.89 kb的YM6552仅获得了7.1%的转染率,EGFP质粒则获得了43.4%的转染效率。在单克隆形成上,较小的pYN3600也获得了更高的单克隆形成数(25个),其中表达EGFP的单克隆有14个,植酸酶PCR阳性集落有11个,高于YM6552的单克隆数(19、8和6)。转基因细胞构建重构胚胎后,所有的胚胎均能表达绿色荧光蛋白,虽其体外发育能力有所下降,但差异不显著(P>0.05)。综上所述,本研究所采用的植酸酶质粒、细胞筛选方法和核移植技术可生产植酸酶重构胚。