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.     

Prevalence of parasite eggs and cysts in faeces from dairy cows in Colombia

Summary When 1, 711 bovine faecal samples from 113 farms in eight dairy areas of Colombia were examined for the presence of helminth eggsFasciola hepatica eggs were found in the faeces from 60% of the farms and samples from animals kept above 2,000 m. Strongyle eggs were found in faeces from 82% of the farms and in 18% of the samples.

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Resumen Se examinaron 1.711 muestras fecales provenientes de 113 fincas, localizadas en ocho zonas lecheras de Colombia. Se encontraron huevos deFasciola hapatica, en el 60% de fincas y muestras fecales de animales sobre los 2,000 m.s.n.m. Se encontraron tambien huevos de estrogilideos en heces del 82% de fincas y en el 18% de las muestras examinadas.

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Résumé La recherche d’oeufs d’helminthes sur 1711 échantillons de fécès provenant de 113 fermes de 8 districts laitiers de Colombie a permis de noter la présence d’oeufs deFasciola hepatica dans 60 p. 100 des fermes et échantillons originaires d’animaux vivant à plus de 2,000 m. Des oeufs de strongles ont été notés dans les fécès provenant de 82 p. 100 des fermes et 18 p. 100 des échantillons.

     

A method for the interpretation of parasite egg counts in faeces of sheep

A new method for the interpretation of strongyloid egg counts in faeces in sheep flocks is proposed. The "Flock Parasitism Index" (FPI) is based on three epizootiological concepts: extent of parasitism, distribution of its intensity, and risk of pasture contamination by eggs. In the estimation of FPI from a number of faecal samples from different animals in a flock, the percentage of parasitized sheep and the distribution of parasitized animals of the sample into the classical categories of "low", "moderate" and "high" from the number of eggs per gram of faeces (EPG) and the arithmetic mean of EPG of the sample are considered. The pathogenic and epizootiological importance of each one of the "low", "moderate" and "high" categories are considered to be 0.1, 0.5 and 1, respectively. From the relationship between EPG and worm burdens, probabilities for "low", "moderate" and "high" distributions are calculated, and from the percentages of worm burdens estimated and their levels of confidence at 95% it is suggested that values of FPI equal to 2.0 must be considered as the limit of parasitism where economic losses are tolerable.     

Comparison of gastrointestinal parasite communities in vervet monkeys

Globally, habitat degradation is accelerating, especially in the tropics. Changes to interface habitats can increase environmental overlap among nonhuman primates, people, and domestic animals and change stress levels in wildlife, leading to changes in their risk of parasite infections. However, the direction and consequences of these changes are unclear, since animals may benefit by exploiting human resources (e.g., improving nutritional health by eating nutritious crops) and decreasing susceptibility to infection, or interactions with humans may lead to chronic stress and increased susceptibility to infection. Vervet monkeys are an excellent model to understand parasitic disease transmission because of their tolerance to anthropogenic disturbance. Here we quantify the gastrointestinal parasites of a group of vervet monkeys (Chlorocebus aethiops) near Lake Nabugabo, Uganda, that frequently overlaps with people in their use of a highly modified environment. We compare the parasites found in this population to seven other sites where vervet monkey gastrointestinal parasites have been identified. The vervets of Lake Nabugabo have the greatest richness of parasites documented to date. We discuss how this may reflect differences in sampling intensity or differences in the types of habitat where vervet parasites have been sampled.     

Embryonation and infectivity of Ascaris suum eggs. A comparison of eggs collected from worm uteri with eggs isolated from pig faeces

Ascaris suum eggs were collected from pig faeces or dissected from worms obtained from the same pigs. Eggs from the two sources were allowed to embryonate in 0.1 N H2SO4, in 1% buffered formalin or in tap water. The embryonation of the sulphuric acid and water cultures occurred at the same speed, while the formalin cultures developed slightly more slowly. By experimental inoculation of helminth-free pigs and subsequent counting of white spots in the livers and larvae in the lungs day 7 p.i., the infectivity of eggs dissected from worm uteri and embryonated in sulphuric acid (a normal laboratory procedure) was compared with that of eggs collected from faeces and embryonated in water (i.e. more naturally developed eggs). The results suggest that the two types of eggs were equally infective. For this reason the common practice of using Ascaris eggs dissected from worms for experimental infections might be acceptable.     

Use of PCR-RFLP to identify Leishmania species in naturally-infected dogs

Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania (Leishmania) chagasi and Leishmania (Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive (≥1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic (n = 12) and asymptomatic (n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. (L.) chagasi except one, infected with L. (V.) braziliensis. PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides.     

Optimization of the procedure for counting the eggs of Fasciola gigantica in bovine faeces

This paper describes a method for counting eggs of F. gigantica in bovine faeces that optimizes the proportion of eggs recovered and the repeatability of estimates. The method uses 3 g of faeces suspended in 0.05% Tween 20. The suspension is passed through three 6 cm diameter sieves in tandem to remove fibrous debris, with respective apertures of 1 mm, 450 μm, and either 266 or 200 μm. The filtrate is allowed to sediment for 3 min in a conical flask; the sediment is recovered, then resuspended in 200 ml of 0.05% Tween 20 and allowed to sediment. After 3 min the sediment is washed in a sieve with an aperture of 53 μm, which retains the eggs. Eggs suspended in 15 ml of 1% methylene blue are counted using a dissecting microscope. Use of Tween 20 instead of water as the suspending agent for faeces gave a significant threefold increased the proportion of eggs recovered and reduced variability between repeated counts. This method is able to detect about one-third of the eggs present. It was concluded that the high proportion of F. gigantica eggs lost may be due to the presence of hydrophobic and covalent bonds on the eggs that bind them to debris, with which they are discarded.     

Evaluation of techniques for detection of small trematode eggs in faeces of domestic animals

This study was conducted to evaluate techniques for detection of small trematode eggs in faeces of dogs, cats and pigs. Faecal samples from dogs (n=80), cats (n=35) and pigs (n=114) were examined by Kato-Katz technique (KK), formalin-ether sedimentation technique (FE) and a method of combining: filtration, sedimentation and centrifugation, developed at DBL - Centre for Health Research and Development (former Danish Bilharziasis Laboratory) (DBL). Necropsy was performed on 38 dogs, 25 cats and 16 pigs and was considered as a gold standard method for evaluation of the techniques. The results showed 100% specificity for the three techniques. Lower sensitivity was seen for the KK-technique in dog samples in comparison to that for DBL- and FE-technique. The sensitivity of the three techniques was similar in cats and pigs. Based on these findings and practical issues, DBL-technique was chosen as most suitable because the eggs were easily detected and quantified. No toxic chemicals or special equipment were required in comparison with FE-technique that needs ether solution and thus fume cupboards which are often unavailable in local veterinary centres in Vietnam and other developing countries.     

An assessment of a composite sampling method for counting strongyle eggs in sheep faeces

The conventional method for estimating the average strongyle egg count for a group of sheep was compared with a single count from a group composite faecal sample. Sixty-one groups of field samples were used. Composite samples were prepared in the laboratory by pooling equal amounts of faeces from individual samples. Data were logtransformed for analysis to meet the assumption of normality. There were no significant differences in the variances and overall mean counts obtained by the 2 methods. The regression line of log (composite count) on log (group arithmetic mean) did not differ significantly from the line of identity. When untransformed egg count data were categorised as low, moderate and high, the 2 methods were in agreement for 53 of the 61 groups. The mixing and counting process used for both methods (modified McMaster technique) gave highly repeatable results (repeatability = 0.94). The composite method was a quicker and valid alternative to the conventional method for monitoring helminthosis in sheep flocks.     

固原地区市售新鲜蔬菜寄生虫虫卵污染情况调查

目的：了解固原地区市售新鲜蔬菜寄生虫卵污染情况。方法：随机抽样分布于固原地区（黑城、彭阳、海原、红河、李寨）5个村镇菜市场的蔬菜,每个菜市场采购10种蔬菜（以凉拌蔬菜为主）各500 g,每种10份,共计500份,随意分发到当地农家,农家主妇第1次洗涤,笔者第2次洗涤,收集洗菜水各500 mL,分别标记。取沉淀物,用饱和盐水浮聚法制片,显微镜下观察,不论虫种检出一个虫卵或幼虫判为阳性,数据经统计学分析。结果：检查10种蔬菜,发现10种有蛔虫卵,9种有蛲虫卵,6种有钩虫卵,4种有绦虫卵,2种有鞭虫卵。500份标本中,2次洗涤后都检出了寄生虫卵（P〈0.05）,第1次洗涤检出率为23.82%,甘蓝最高,为32.86%,辣椒最低,为14.29%;第2次洗涤检出率为10.04%。还是甘蓝最高,为15.71%,西红柿最低,为5.71%。菜洗的次数越多,虫卵的种类越少,数量也越少。结论：固原市市售蔬菜寄生虫卵总体检出率较高,居民生吃蔬菜有感染寄生虫的机会,应注意提高防病意识。     

Quality characteristics of quail eggs

1. An experiment was carried out with laying Japanese quail to study the influence of age on their egg quality.

2. Egg, yolk, albumen and shell plus membrane weights increased, whereas the egg shape, specific gravity and shell thickness decreased between 49 and 154 days of age.

3. Yolk, albumen and shell plus membrane comprised 32–59%, 53–57% and 7–80%, respectively of the total egg weight.

4. Regression between the quail's age and the characteristics of eggs were derived and correlation coefficients ranged from —0–42 to — 0–86. The albumen weight was more highly correlated with egg weight than yolk weight.

5. Relationships were also derived between egg shape and specific gravity, and egg shape and albumen weight.

6. It was concluded that the eggs’ characteristics were modified as the quails aged, and the best criterion of the shell's strength was the egg shape.     

Use of BAER to identify loss of auditory function in older horses

Objective To use the brainstem auditory evoked response (BAER) to test the hypothesis that auditory function could be worse in older horses than in younger horses. Procedure BAER waveforms in response to click stimuli were measured in five younger horses (5–8 years) and four older horses (17–22 years). Results Compared with the younger horses, the older horses showed significantly (P < 0.02) worse BAER thresholds and significantly (P < 0.02) worse BAER wave V amplitudes to the 90 decibels above normal hearing level stimulus. These results were consistent with partial deafness in the older horse group. Conclusion BAER assessment can be used to identify partial deafness in older horses. Such horses should be managed appropriately, with particular care taken in noisy environments where hearing loss could put the horse and/or its owner at risk of harm.     

Use of alpha-naphthyl acetate esterase staining to identify T lymphocytes in cattle

Peripheral blood lymphocytes (PBL), lymph nodes, and/or spleens from clinically normal cattle were examined for cytochemical staining of alpha-naphthyl acetate esterase (ANAE). Two types of positive-staining patterns in ANAE staining resulted. By a combination of ANAE staining and latex-ingesting test, diffuse ANAE-positive cells were considered as mononuclear phagocytic cells. Using erythrocyte rosettes, erythrocyte antibody complement rosettes, nylon-wool column technique, surface immunoglobulin (SIg) staining, and the ANAE staining technique, granular ANAE-positive lymphocytes were shown to be T lymphocytes. The frequency of T and B lymphocytes in PBL, spleens, and lymph nodes of clinically normal cattle was measured, using ANAE staining and SIg staining. In PBL, 47.7% were ANAE-positive and 26.9% were SIg-positive; in spleens, 22.4% were ANAE-positive and 53.7% were SIg-positive; and in lymph nodes, 38.5% were ANAE-positive and 28.3% were SIg-positive. The frequencies of T and B lymphocytes in PBL, spleens, and/or lymph nodes from cattle with enzootic bovine leukosis (EBL) and cattle with persistent lymphocytosis and in tumor cells from cattle with EBL were measured. When compared with those of clinically normal cattle, PBL, spleens, and lymph nodes of cattle with EBL and the PBL of cattle with persistent lymphocytosis contained numerous SIg-positive cells and few ANAE-positive cells. Tumor cells from cattle with EBL contained 7.3% ANAE-positive and 78.0% SIg-positive cells.