Combined Molecular and Lectin Binding Assays to Identify Different Trichostrongyle Eggs in Feces of Sheep and Goats from Egypt

Triple lectin staining of trichostrongyle eggs from naturally infected small ruminants

The binding of a panel of 19 lectins to carbohydrates on the eggs of economically important nematode parasites of sheep has been assessed as the basis of a rapid test to distinguish parasite eggs, at least at the genus level. A total of six lectins can be used to identify eggs of six nematode parasites: peanut agglutinin (PNA) for Haemonchus contortus; Lens culinaris agglutinin (LCA) for Teladorsagia sp; Aleuria aurantia agglutinin (AAL) for Trichostrongylus sp; Psophocarpus tetragonolobus‑II (PTLII) for Nematodirus sp; Lotus tetragonolobus lectin (LTL) for Cooperia sp and wheat germ agglutinin (WGA) for Chabertia ovina. For WGA, LCA and LTL, weak binding was also observed to H. contortus and Teladorsagia sp, Trichostrongylus sp and C. ovina eggs, respectively. Nematode eggs in two faecal samples were identically identified by both lectin binding and PCR, except for PCR identification of the eggs of Nematodirus sp, as these did not lyse. Lectins bound best to H. contortus eggs extracted from fresh faecal samples or after storage at room temperature or 4°C for up to 24h, but eggs stored at −20°C or −80°C did not bind PNA.

Lectin binding to carbohydrates on parasite surfaces has been investigated as a method of distinguishing adult worms, eggs and sheathed and exsheathed L3 of Teladorsagia circumcincta and Haemonchus contortus, economically important abomasal parasites in temperate climates. Both species were maintained as pure laboratory cultures of field isolates from New Zealand. Each of the four life cycle stages could be distinguished by the binding of at least one lectin: adult worms by Sambucus nigra agglutinin (SNA); eggs by peanut agglutinin (PNA), ConcavalinA and Lens culinaris agglutinin (LCA); exsheathed L3 by Griffonia simplicifolia-I lectin (GSL-I) and Lotus tetragonolobus lectin (LTL) and sheathed L3 by Aleuria aurantia lectin (AAL). The whole surface of both adult T. circumcincta and H. contortus strongly bound lectins specific for N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), mannose and fucose, but the two species could be distinguished by SNA binding only to T. circumcincta. Eggs could be distinguished by the binding of mannose-specific PNA to H. contortus and GalNAc-specific LCA and PSA to T. circumcincta eggs. GalNAc, GlcNAc and mannose lectins bound to the cuticle and over the excretory pores of a large proportion of sheathed L3 of both species, but only the H. contortus surface had exposed fucose or sialic acid complexes. The distinguishing lectin for sheathed L3 was AAL, which did not bind to T. circumcincta, but bound weakly to the head region of all fresh H. contortus and to 50-90% after 3months storage. The cuticle of exsheathed L3 was unresponsive to all 19 lectins, and any binding was restricted to the head and tail regions. L3 exsheathed after 2-4months storage could be distinguished by the binding of GSL-I and LTL to H. contortus but not to T. circumcincta. Lectin binding could be a useful adjunct in identifying L3, but lacked the consistency to be definitive, whereas it could be further developed as a practical method of distinguishing parasitic nematodes at other stages in the life cycle, particularly the eggs.

Molecular characterization of Cysticercus tenuicollis of slaughtered livestock in Upper Egypt governorates

This paper reports the use of near infrared and mid-infrared spectroscopy to detect the presence and quantity of eggs of the gastrointestinal nematode Haemonchus contortus in sheep faeces. Haemonchus contortus eggs were quantified in dried, finely ground sheep faeces and in moist, coarsely ground faeces using near infrared and mid-infrared bench top spectrometers and a portable near infrared spectrometer. When Haemonchus contortus eggs were presented without faecal medium, it was found that the wavelength region of 1880–2100 nm was most important for detection. Broad classes of chemical properties found in the near infrared region were identified for dried Haemonchus contortus eggs using a mid-infrared spectrometer. However, when Haemonchus contortus eggs were mixed into the complex matrix of sheep faeces, the development of a robust calibration model for egg detection proved to be challenging (R2 < 0.47). The reliability of this method, if used for the detection of Haemonchus contortus eggs in the field, may be further limited by variations in egg species, faecal moisture content, faecal composition and particle size. Nevertheless, this is the first report identifying near infrared bands for Haemonchus contortus eggs and provides valuable information for future studies towards a spectroscopy-based method for detection of gastrointestinal nematodes.

Some problems in fiber determination of a tannin-rich forage ( Acacia saligna leaves) and their implications in in vivo studies

Seroepidemiological study of Toxoplama gondii in small ruminants (sheep and goat) in different provinces of Mongolia

TRANSMISSIBLE spongiform encephalopathies (TSEs), or prion diseases, are a group of fatal neurodegenerative diseases that comprise scrapie in sheep and goats, bovine spongiform encephalopathy (BSE) in cattle and Creutzfeldt-Jakob disease...

In recent years, the consumption of milk and dairy products has dramatically increased in several parts of the world. Different livestock plays an essential role in global milk production. This study was designed to evaluate different chemical and physical components of milk in four groups of livestock, including cows, buffalos, sheep, and goats. To this end, 200 raw milk samples were collected from cows, buffalos, sheep, and goats (n=50) across Dhi-Qar Governorate, Iraq, for a period of one year (from 01.10.2018 to 01.06.2019). The findings showed sheep and buffalos' milk samples had a significantly higher percentage of total solids (TS%), compared to cows and goats' milk samples (P<0.05). However, there were no significant differences in the TS% between sheep and buffalos' milk samples. Furthermore, the mean TS% values in cows, buffalos, sheep, and goats' milk samples were determined at 11.14%, 12.87%, 13.26%, and 11.33%, respectively. As for fat percentage (F%), buffalos' milk samples had significantly higher F% (4.80%), compared to milk samples of cows, sheep, and goats (P<0.05). Additionally, sheep's milk samples had significantly higher F% (P<0.05) than cows and goats' milk samples determined at 2.78%, 4.20%, and 2.98%, respectively. The findings showed the percentage of solids not fat (SNF%) was significantly higher in sheep's milk (8.97%), compared to milk samples of cows, buffalos, and goats (P<0.05). Additionally, it was found that the SNF% was significantly higher (P<0.05) in Buffalos' milk samples, compared to cows and goats' milk samples determined at 8.36%, 8.60%, and 8.35%, respectively. Moreover, the results revealed that the percentage of milk protein content in sheep's milk was significantly higher than the cows, buffalos, and goats' milk (P<0.05). Recorded data also showed no significant differences in the percentage of milk lactose among cows, buffalos, sheep, and goats' milk samples (P<0.05). Furthermore, the findings illustrated that the percentage of milk ash (Ash%) in sheep's milk samples was significantly higher than the cows, buffalos, as well as goats' milk samples (P<0.05), and no significant differences were observed among cows, buffalos, and goats' milk samples in the Ash% (P<0.05). In addition, there were no significant differences in the specific gravity among different milk samples (P<0.05). Finally, the results displayed no significant differences between cows and goats' milk samples in all the studied traits (P<0.05).

The effects of arsenic contaminated drinking water of livestock on its total levels in milk samples of different cattle: Risk assessment in children

Serological evidence for the presence of influenza D virus in small ruminants

Prevalence of gastrointestinal parasites in small ruminants in Grenada, West Indies.

Molecular detection of the B1 gene of Toxoplasma gondii in blood samples of female sheep and goats in Tebessa, northeastern Algeria

Toxoplasmosis is one of the most common meat‐borne parasitic infections worldwide. Consumption of raw or undercooked meat which containsToxoplasma gondiitissue cysts is an important route of human infection. In this study, we investigate the serological and molecular prevalence ofT. gondiiinfection in sheep and goat samples in Kashan, Iran, from 2015 to 2016. Serological (IgG antibody) and molecular detections were performed by the enzyme‐linked immunosorbent assay and polymerase chain reaction on the sera and heart samples of sheep (n = 90) and goats (n = 90), respectively.T. gondii‐IgG antibody was detected in 12.2% of sheep and 4.4% of goat samples. The parasite's DNA was detected in 17.8 and 8.9% of sheep and goat samples, respectively.Practical applicationsThe results of this study emphasize on the role of the sheep and goat as reservoirs ofT. gondiiinfection. Hence, consumption of adequately cooked meat should be considered for prevention ofT. gondiiinfection in humans.

Glycoproteins have many important biological functions. In particular, aberrant glycosylation has been observed in various cancers, such as liver cancer. A well-known glycoprotein biomarker is α-fetoprotein (AFP), a surveillance biomarker for hepatocellular carcinoma (HCC) that contains a glycosylation site at asparagine 251. The low diagnostic sensitivity of AFP led researchers to focus on AFP-L3, which has the same sequence as conventional AFP but contains a fucosylated glycan. AFP-L3 has high affinity for Lens culinaris agglutinin (LCA) lectin, prompting many groups to use it for detecting AFP-L3. However, a few studies have identified more effective lectins for fractionating AFP-L3. In this study, we compared the amounts of enriched AFP-L3 with five fucose-specific lectins─LCA, Lotus tetragonolobus lectin (LTL), Ulex europaeus agglutinin I (UEA I), Aleuria aurantia lectin (AAL), and Aspergillus oryzae lectin (AOL)─to identify better lectins and improve HCC diagnostic assays using mass spectrometry (MS). Our results indicate that LTL was the most effective lectin for capturing AFP-L3 species, yielding approximately 3-fold more AFP-L3 than LCA from the same pool of HCC serum samples. Thus, we recommend the use of LTL for AFP-L3 assays, given its potential to improve the diagnostic sensitivity in patients having limited results by conventional LCA assay. The MS data have been deposited to the PeptideAtlas (PASS01752).