Sero-epidemiology of peste des petits ruminants (PPR) in Pakistan

A sero survey was conducted during 2005-2006 to estimate the sero prevalence of PPR in the small ruminant population of Pakistan. A total of 2798 samples were collected including goats (1979) and sheep (819) from villages in 27 randomly selected districts. These were tested by cELISA for PPRV and true prevalence estimates were calculated by Rogan and Gladen estimator. Overall, 1273 (45.5%) were found positive; 980 (49.5%) of 1979 samples from goats and 293 (35.8%) of 819 serum samples from sheep were positive. The true sero-prevalence of PPR was estimated to be 48.5% (95% CI, 46.6-50.3), and 52.9% (95% CI, 50.7-55.1) and 37.7 (95% CI, 34.4-41.0) for goats and sheep, respectively. PPR virus is widely distributed all across Pakistan and has become an endemic infection of small ruminants. Since it is one of the leading causes of morbidity and mortality in small ruminants, it poses a serious threat to food security and the rural economy in Pakistan.     

Experimental infection of alpine goats with a Moroccan strain of peste des petits ruminants virus (PPRV)

Peste des petits ruminants virus (PPRV) recently caused a serious outbreak of disease in Moroccan sheep and goats. Alpine goats were highly susceptible to PPRV with mortality rates approaching 100%, as opposed to local breeds of sheep which were less susceptible to the disease. The relative susceptibility of alpine goats was investigated through an experimental infection study with the Moroccan strain of PPRV. Severe clinical signs were observed in the alpine goats with virus being excreted through ocular, nasal and oral routes. No difference in the severity of the disease in goats was observed with different inoculation routes and transmission of the virus by direct contact was confirmed. This study confirmed the susceptibility of the alpine goat to PPRV infection and describes a challenge protocol that effectively and consistently reproduced severe clinical signs of PPR in experimentally infected goats.     

小反刍兽疫病毒重组N蛋白抗原制备及间接ELISA检测方法的建立

将编码小反刍兽疫病毒(PPRV)N蛋白的基因全长克隆到pBacPAK9载体中,并在昆虫细胞中进行表达。对表达产物进行SDS-PAGE分析,并用PPRV阳性血清进行了Western blot鉴定。用Ni-IDA亲和柱对组氨酸融合表达的N蛋白进行了纯化,并对纯化产物进行了理化性质和抗原活性分析。最后以表达的N蛋白作为包被抗原,建立间接ELISA检测方法,临床检测137份山羊血清并与IDvet公司的商品化PPRV抗体检测试剂盒进行比较。结果表明,制备的PPRV N蛋白抗原在溶液中以单体形式存在,具有非常好的抗原活性。用该蛋白建立的间接ELISA检测方法与IDvet试剂盒符合率为956%。本研究为小反刍兽疫病毒重组N蛋白抗原制备相关质量标准的建立奠定了基础,同时建立的ELISA抗体检测方法可以很好地用于小反刍兽疫的临床诊断。     

<Emphasis Type="Italic">Theileria</Emphasis> (<Emphasis Type="Italic">Babesia</Emphasis>) <Emphasis Type="Italic">equi</Emphasis> and <Emphasis Type="Italic">Babesia caballi</Emphasis> Infections in Horses in Galicia,Spain

The control of equine piroplasmosis is becoming increasingly important to maintain the international market open to the horse industry. The purpose of this study was to demonstrate the occurrence of equine piroplasmosis (Theileria equi and Babesia caballi) in Galicia, north-west Spain, and to compare haematological and serum biochemistry parameters between non-parasitaemic horses and horses parasitaemic with T. equi and B. caballi. Sixty serum samples (control group) were taken from healthy horses pastured on two farms, and examined for evidence of equine T. equi and B. caballi infection by indirect fluorescent antibody test (IFAT). Of the 60 samples, 24 (40%) and 17 (28.3%) samples were positive for T. equi and B. caballi, respectively. Twelve (20%) samples were positive for both parasites. Haematology and serum biochemistry were compared between controls and a series of 36 horses clinically affected by T. equi (25) or B. caballi (11). Compared with the healthy group, there was a 43% and 37% decrease in the haematocrit for T. equi and B. caballi infection, respectively. Parasitaemic horses presented an intense anaemia and serum biochemistry signs of liver damage. The anaemia was more severe in T. equi-infected than in B. caballi-infected horses. Our results suggest that equine piroplasmosis is widespread in the region and is a cause for concern.     

Seroprevalence of <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> infection in yaks (<Emphasis Type="Italic">Bos grunniens</Emphasis>) in northwestern China

The seroprevalence of Toxoplasma gondii infection in yaks (Bos grunniens) was surveyed in Qinghai Province, northwestern China in May and June 2010. A total of 650 serum samples were collected from six counties and assayed for T. gondii antibodies by an indirect hemagglutination test. Antibodies to T. gondii were found in 35.08% (228/650) with the highest rate of 55.34% in Chengduo County. The results of the present survey indicated that infection with T. gondii in cattle is widely spread in China, including yaks in Qinghai Province.     

Experimental infection of <Emphasis Type="Italic">Peste des Petit Ruminant</Emphasis> virus and <Emphasis Type="Italic">Mannheimia haemolytica</Emphasis> A2 in goats: immunolocalisation of <Emphasis Type="Italic">Mannheimia haemolytica</Emphasis> antigens

Nigerian strain of Peste des Petit Ruminant (PPR) virus and Mannheimia haemolytica (MH) biotype A serotype 2, was used successfully to reproduce a concurrent disease in West African Dwarf goats. The development of the various pathological features were studied at regular intervals following infection. The acute inflammatory reaction which had developed by day 3 after initial infection was characterised by flooding of the alveoli by neutrophils, oedema, hemorrhage and syncytial cells together with a moderate bronchial and bronchiolar epithelial necrosis. This progressed to a milder acute broncho interstitial pneumonia with giant cells. At this stage, the mucosal immunity were well developed especially the aggregate form of NALT and more of nodular forms of BALT. The organisms were demonstrated with strong immunostaining in the necrotic center, necrotic alveolar wall, fibrin, serous exudate, and degenerated leukocyte in the alveoli and respiratory airways. The bacterial antigens were observed as a strong immunostaining in the blood vessels of the nasal septum, sinusoid in the liver and interstium of the kidney, cytoplasm of alveolar macrophages, pneumocytes, bronchial and bronchiolar epithelium, in the monocytes in the blood vessels. These findings confirmed the enhancement of MH tropism especially in the respiratory tract, liver and kidney. It also showed that West african dwarf goats are highly susceptible to the intratracheal combined infection of PPR virus and MH. The fact that the infection induces strong mucosal responses, this phenomenon can be explored in Africa with the use of combined PPR virus and MH intranasal vaccines to curtail the menace of pneumonia associated with the combined infection on field.     

Prevalence and molecular diagnosis of <Emphasis Type="Italic">Trypanosoma evansi</Emphasis> in Nili-Ravi buffalo (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>) in different districts of Punjab (Pakistan)

The prevalence of Trypanosoma evansi was investigated in 1,250 Nili-Ravi buffaloes of mixed age and sex by polymerase chain reaction (PCR) for the first time in Pakistan. DNA of the trypanosomes was isolated with TRIREAGENT®. The assay was employed using primers ESAG 6/7, specific for a 237-bp fragment from T. evansi genomic DNA. The samples were screened for the presence of T. evansi also by stained thin smear. Forty-four (3.5%) samples were positive by microscopy, while 97 (7.7%) samples were identified by PCR, indicating the high sensitivity of PCR for surveying the disease in epidemiological studies.     

Studies on the development of omasum in west african dwarf goats (<Emphasis Type="Italic">Capra hircus</Emphasis>)

This work studied the sequential morphological changes of omasum in foetuses, neonates and adult West African Dwarf (WAD) goats by gross and light microscopic methods. The mean omasal volume was 1.37 ± 0.36, 4.2 ± 0.4, 8.7 ± 6.9 and 60.1 ± 8.6 ml for gestation day 87 and 146 foetuses, neonates and adults respectively. Grossly the mucosa exhibited longitudinally oriented primary, secondary, tertiary and quartnery laminae with smooth surface in foetuses and numerous papillae in the adults. Microscopically it was lined by stratified squamous epithelium which was divided into larger lighter luminal and smaller darker basal zones in gestation day 60 foetuses. The core of the laminae contained extension of the inner muscular tunic. The basal zone developed lateral evaginations (corial papillae) into the lighter zone of the laminae in gestation day 106 foetuses. By term these corial papillae approached the luminal surface. The papillae emerged above the luminal surface in 4 week old neonates. The papillae were fully developed in adult goats. The mean papillary height and width were -205.1 ± 34.0, 67.0 ± 9.6; 235.0 ± 86.5, 185.0 ± 42.3 and 570.3 ± 60.0, 290.1 ± 66.3 μm for foetuses, neonates and adult goats. The inner circular layer was thicker than the outer longitudinal layer of the muscular tunic. The central muscular layer of the laminae originated from the inner circular layer with attachment to the outer muscular layer in adults. This attachment could be adaptation for better anchorage and muscular contraction of the laminae in WAD goats.     

Seasonal variations in plasma glucocorticoid levels in yaks (<Emphasis Type="Italic">Poephagus grunniens L</Emphasis>.) (<Emphasis Type="Italic">Bos grunniens</Emphasis>)

The changes of glucocorticoid concentrations were measured by highly sensitive enzymeimmunoassay procedures using second antibody coating technique and cortisol -HRP amplified system for one whole calendar year in yaks. The year was divided into seasons of cool, intermediate and hot seasons. The glucocorticoid values were lower (P < .01) in hotter months than in cool and intermediate seasons. Rectal temperature and respiration rates were indicative of heat stress in the hot temperature season and gave significant negative correlation with circulating corticoid concentration.     

Evidence of mixed infection of peste des petits ruminants virus and bluetongue virus in a flock of goats as confirmed by detection of antigen, antibody and nucleic acid of both the viruses

A mixed infection with peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) occurred in goats which exhibited symptoms characteristic of PPR. A number of samples were collected from ailing or dead goats for labrotory diagnosis. Antibody to BTV and PPRV was detected in sera samples by competitive ELISA. No PPRV antigen was detected in tissue samples like lung and spleen, however, presence of PPRV antigen in some sera samples was confirmed by sandwich ELISA. All the blood samples collected from the ailing animals were found positive for BTV antigen by a sandwich ELISA. BTV- and PPRV nucleic acids were amplified from the pooled blood and tissue samples respectively by RT-PCR assays. The identity of the amplicons was confirmed by cloning and sequencing. All these tests confirm that the goats were infected with PPRV and BTV simultaneously. Isolation of viruses from the clinical samples is underway.     

Prevalence of thin-walled <Emphasis Type="Italic">Sarcocystis cruzi</Emphasis> and thick-walled <Emphasis Type="Italic">Sarcocystis hirsuta</Emphasis> or <Emphasis Type="Italic">Sarcocystis hominis</Emphasis> from cattle in Iran

Bovine sarcocystosis is caused by Sarcocystis cruzi and is known to cause considerable morbidity and mortality in cattle. This species is distributed worldwide in cattle and is the most prevalent of the Sarcocystis species infecting cattle. There is high infection rate of sarcocyst in cattle in Iran, but to our knowledge, there is no study about identification of Sarcocystis species. This work aimed to survey prevalence of S. cruzi cyst in slaughtered cattle of Isfahan, Iran. In this study, esophageal and diaphragmatic muscles of 100 cattle were collected from Fesaran abattoir of Isfahan and examined for the presence of Sarcocystis spp. cysts macroscopically and microscopically. No macroscopic sarcocysts were found in any of the samples. In light microscopy, 89 out of 100 cattle (89%) had thin-walled cysts of S. cruzi, while 21 out of them (21%) had thick-walled sarcocysts. In addition to light microscopy, ultrastructural features of the thin-walled cyst confirmed the presence of S. cruzi.     

Ocular infection of donkeys (Equus asinus) with <Emphasis Type="Italic">Setaria equina</Emphasis>

Seven donkeys raised in different locations in Egypt were found to have a unilateral eye showing motile white worms in the aqueous humor. The parasites were surgically removed from the anterior chamber of the eye in five out of the seven donkeys, and were cleared, mounted and identified as Setaria equina based on light microscopic features. The ocular infection with S. equina reported herein may be the first reported aberrant cases in the Egyptian animals.     

Comparative study of experimental Foot-and-Mouth Disease in cattle (<Emphasis Type="Italic">Bos indicus</Emphasis>) and buffaloes (<Emphasis Type="Italic">Bubalis bubalus</Emphasis>)

Foot-and-mouth disease (FMD) is one of the most contagious diseases affecting wide range of host species with variable severity and decreased productivity. The present study was undertaken to compare the clinical and leucocytic changes in indigenous Indian cattle and buffaloes experimentally infected with FMD virus (FMDV) Asia 1. A mild type of disease was observed in the cattle, more so in buffaloes infected with FMDV. Difference in terms of type, site and healing of lesion was observed between cattle and buffaloes. Foot lesions were more common than tongue in buffaloes, which were mainly evident in bulb of the heel in contrast to interdigital foot lesions in cattle. Further, FMDV infection induced a transient moderate leucopenia with lymphopenia in both cattle and buffaloes, but monocyte levels diverged. Relationship between the raised body temperature, leucocytic changes and lesion development was observed. Microscopic changes were observed in the keratinized epithelium of tongue and foot. The findings of the present study indicated the need to investigate the early leucocytic changes in cattle and buffaloes in depth for better understanding of the disease process.