Activatable Shiga toxin 2d (Stx2d) in STEC strains isolated from cattle and sheep at slaughter

Shiga toxin producing Escherichia coli (STEC) harbouring the stx(2d-activatable) gene and expressing the mucus- and elastase-activatable phenotype have been associated with severe outcomes of human disease. However, there is limited data available on the occurrence of such strains in livestock reservoirs. In this study, we analyzed 11 STEC strains isolated from healthy cattle and sheep at slaughter that were originally detected to contain the stx(2c) allele, for the presence of the stx(2d-activatable) genotype. Ten of the eleven strains displayed the stx(2d-activatable) genotype as determine by PstI restriction fragment length polymorphism (RFLP) of 890-bp fragments of their stx genes. However, only in 6 of the 10 strains whose stx genes were sequenced, the presence of stx(2d-activatable) could be confirmed based on the predicted amino acid sequence of their StxA subunits; the remaining four strains contained Stx2c A subunit. Five of the six strains which contained stx(2d-activatable) displayed the activatable phenotype on Vero cells. Genes for adhesins such as the outer membrane protein intimin (eae), which is essential for the intimate attachment and the formation of attaching-and-effacing lesions on intestinal epithelial cells, or the STEC autoagglutinating adhesin (saa), potentially important in eae-negative STEC, were not detected. Moreover, all the strains tested negative for EHEC-hlyA encoding enterohaemorrhagic E. coli (EHEC) hemolysin. To our knowledge, this is the first study that reports the presence of STEC harbouring stx(2d-activatable) and producing the activatable Stx2d in fecal samples of sheep. Therefore both cattle and sheep are reservoirs of such strains and potential sources of human infections. This is of particular importance, because in contrast to other eae-negative STEC, strains producing Stx2d(activatable) may cause severe diseases such as bloody diarrhoea and haemolytic uremic syndrome in humans.     

P-80 Clinical aspects of chronic progressive pastern dermatitis in Belgian draft horses

The aim of this study was to evaluate the fungi present on the skin surface in healthy horses with a particular interest in Malassezia on predisposed skin areas. Twenty-five mares were included, each sampled on nine skin areas including those most likely contaminated by Malassezia : nostrils, lips, ear pinnae, base of the mane, pasterns, pectoral areas, lateral thorax, udder and perianal areas. Two sterile carpets were applied to the skin, and then used for inoculation of agar (Sabouraud's plus cycloheximide and chloramphenicol), both with and without lipids for isolation of different species of Malassezia . This method is currently successful in isolating Malassezia in dogs. Cultures were incubated at 32°C, observed on day 5 for Malassezia , and then maintained at 27°C for complementary morphological identifications. From 223 cultures on standard medium, only seven remained sterile. The most common fungi were filamentous (number of positive samples): Scopulariopsis (156), Penicillium (79), Acremonium (67), Aspergillus (53), Paecilomyces spp. (38), and Stachybotrys (33). Interestingly, Microsporum gypseum was also isolated (14 samples from seven areas) from 10 horses. Yeast or yeast-like fungi were Geotrichum (52), Trichosporon (21), and an additional 33 samples being positive for yeast other than Malassezia . Results from the other 223 cultures on lipid-rich agar were similar. Most of the fungi were isolated from the nine areas selected, but Malassezia was never isolated from 446 cultures.
Funding: Self-funded.     

Bioactivity of formulas containing essential oils from the family Myrtaceae for the management of deltamethrin-resistant Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Plutella xylostella is the major pest of Brassica and is resistant to more than 95 synthetic insecticides, including deltamethrin, which is frequently used in family farming activities in the state of Pernambuco, Brazil. A possible strategy for overcoming such resistance is the use of botanical insecticides containing essential oils. The aim of the present study was to determine the chemical composition of essential oils from Syzygium aromaticum, Melaleuca alternifolia, Melaleuca leucadendra, Eucalyptus citriodora and Eucalyptus globulus and evaluate the effects of these oils as well as selected constituents (β-pinene, α-terpinene, ρ-cymene, limonene, 1,8-cineole, γ-terpinolene, citronellal, terpinen-4-ol, eugenol, β-caryophyllene and E-nerolidol) on different stages of development of two populations of P. xylostella – one susceptible and one resistant to deltamethrin. Chemical analysis by gas chromatography-mass spectrometry revealed eugenol (S. aromaticum), 1,8-cineole (E. globulus), citronellal (E. citriodora), terpinen-4-ol (M. alternifolia) and E-nerolidol (M. leucadendra) to be the major constituents. Among the oils tested, S. aromaticum was the most promising in the different modes of action and had the same level of toxicity to the two populations of P. xylostella analyzed. Among the constituents, E-nerolidol and terpinen-4-ol exhibited promising larvicidal action, whereas limonene exhibited promising ovicidal and antifeedant action. None of the oils presented severe phytotoxicity to the host plant. All results were compared to those achieved with deltamethrin and azadirachtin, which were used as positive controls. The deltamethrin-resistant population of P. xylostella did not exhibit pre-adaptive resistance to the oil from S. aromaticum or E-nerolidol, terpinen-4-ol and limonene.

     

Factors influencing short-term parasitoid establishment and efficacy for the biological control of Halyomorpha halys with the samurai wasp Trissolcus japonicus

BACKGROUND

Classical biological control has been identified as the most promising approach to limit the impact of the invasive pest species Halyomorpha halys (Heteroptera: Pentatomidae). This study investigated the parasitism rate at sites where the biocontrol agent Trissolcus japonicus (Hymenoptera: Scelionidae) was released and where its unintentional introduction took place, in the Trentino–South Tyrol region. The effect of land-use composition was studied to understand which factors favor the establishment of hosts and parasitoids, including native and exotic species.

RESULTS

The released T. japonicus were detected a year after the start of the program, with a significant parasitoid impact and discovery, compared to control sites. Trissolcus japonicus was the most abundant H. halys parasitoid, and Trissolcus mitsukurii and Anastatus bifasciatus were recorded also. The efficacy of T. mitsukurii was lower in sites where T. japonicus was successfully established, suggesting a possible competitive interaction. Parasitism level by T. japonicus at the release sites was 12.5% in 2020 and 16.4% in 2021. The combined effect of predation and parasitization increased H. halys mortality up to 50% at the release sites. Landscape composition analysis showed that both H. halys and T. japonicus were more likely to be found at sites with lower altitude and with permanent crops, whereas other hosts and parasitoids preferred different conditions.

CONCLUSION

Trissolcus japonicus showed a promising impact on H. halys, at release and adventive sites, with minor nontarget effects, mediated by landscape heterogeneity. The prevalence of T. japonicus in landscapes with permanent crops could support IPM in the future. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Antibodies to elastin peptides in sera of Warmblood horses at different ages

REASONS FOR PERFORMING STUDY: Early diagnosis and monitoring progression of chronic diseases in elastin-rich tissues, such as chronic progressive lymphoedema in draught horses and chronic obstructive pulmonary disease (COPD) is still a real challenge in the horse. Use of an enzyme-linked immunosorbent assay (ELISA) to detect anti-elastin antibody (AEAb) levels might be useful to assess the status of such diseases. Baseline levels, representing physiological breakdown of elastin in normal horses, are not available at present. HYPOTHESIS: Levels of AEAb in healthy horses are generally low and follow the same age-related pattern as found in man. Therefore, elevation of AEAb levels in serum can be used to evaluate pathological elastin breakdown in elastin-rich tissues. METHODS: Sera of 84 clinically healthy Warmblood horses were evaluated for the presence of AEAbs by means of a modified version of an ELISA technique used in man. The horses were divided in 5 age groups: A) < 4 months; B) 4-23 months; C) 2-3 years; D) 4-10 years; and E) > 11 years. RESULTS: Antibodies to elastin were found in all equine serum samples tested. Their levels were lowest in Group A, low in Groups B and E and highest in animals age 2-10 years. CONCLUSIONS: Measuring AEAbs in serum of horses by an ELISA technique proved to be possible and levels were stable during well-defined life stages. POTENTIAL RELEVANCE: Changes in AEAb levels are expected to be useful for early diagnosis and for monitoring progression of diseases that affect elastin-rich tissues, such as chronic progressive lymphoedema and COPD.     

Antibodies to elastin peptides in sera of Belgian Draught horses with chronic progressive lymphoedema

REASONS FOR PERFORMING STUDY: Chronic progressive lymphoedema (CPL) is a recently recognised disease of the lymphatic system characterised by lesions in the skin of the lower legs in several draught horse breeds, including the Belgian Draught hourse. Clinical signs slowly progress and result in severe disfigurement of the limbs. Ideally, supportive treatment should be started early in the disease process. However early diagnosis and monitoring progression of CPL is still a challenge. HYPOTHESIS: Elastin changes, characterised by morphological alterations as well as increased desmosine levels, in the skin of the distal limbs of horses affected with CPL are probably associated with a marked release of elastin degradation products, which elicit production of circulating anti-elastin antibodies (AEAbs) in the serum. An enzyme-linked immunosorbent assay (ELISA) for detection of serum AEAbs may document elastin breakdown. METHODS: An ELISA technique was used to evaluate levels of AEAbs in sera of 97 affected Belgian Draught horses that were clinically healthy except for possible skin lesions, associated with CPL in their distal limbs. The horses were divided into 5 groups according to the severity of these skin lesions: normal horses (Group 1, n = 36), horses with mild lesions (Group 2, n = 43), horses with moderate lesions (Group 3, n = 8), horses with severe lesions (Group 4, n = 10) and, as a control, healthy Warmblood horses, unaffected by the disease (Group 5, n = 83). RESULTS: Horses with clinical signs of CPL had significantly higher AEAb levels compared to clinically normal Belgian Draught horses and to healthy Warmblood horses. These levels correlated with severity of lesions. CONCLUSIONS: CPL in draught horses is associated with an increase of serum AEAbs. POTENTIAL RELEVANCE: Evaluation of serum levels of AEAbs by ELISA might be a useful diagnostic aid for CPL. Pathological degradation of elastic fibres, resulting in deficient support of the distal lymphatics, is proposed as a contributing factor for CPL in Belgian Draught horses.     

Sensitivity of Chlamydia suis to cathelicidin peptides

Nine Chlamydia suis isolates, obtained from pigs with conjunctivitis, were molecularly characterized by ompA sequencing and their in vitro susceptibility to six cathelicidin peptides (SMAP-29, BAC-7, BMAP-27, BMAP-27, BMAP-28, PG-1, LL-37) determined in cell culture. SMAP-29 was the most active peptide, reducing the intracellular inclusion number by > or =50% at a concentration of 10 microg/ml (3 microM) in six of the nine isolates tested. Three molecularly identical isolates were insensitive at a concentration as high as 80 microg/ml (25 microM). Of the remaining cathelicidin peptides tested, BAC-7 and BMAP-27 were active against six C. suis isolates at a concentration of 80 microg/ml (25 and 26 microM, respectively). Cathelicidins LL-37 and PG-1 did not show any anti-chlamydial activity at 80 microg/ml.     

Evaluation of FOXC2 as a candidate gene for chronic progressive lymphedema in draft horses

Chronic progressive lymphedema (CPL) is a debilitating condition identified in Clydesdales, Shires and Belgian draft horses and results in progressive swelling of the lower legs associated with the development of thick skin folds, ulcerations, fibrosis and marked hyperkeratosis. The result is severe discomfort and recurrent secondary infection, often requiring euthanasia. Due to the delayed onset, many horses are bred prior to diagnosis. CPL has only been documented in three related draft horse breeds, suggesting a genetic cause. Determining the molecular basis would enable owners to test horses prior to breeding and facilitate the elimination of CPL. Mutations in the FOXC2 gene cause a comparable condition in humans, lymphedema-distichiasis. This gene was sequenced in affected and unaffected draft horses and a control horse. Four single nucleotide polymorphisms (SNPs) were identified in unaffected draft horses and the control horse, indicating that they were not associated with CPL. A fifth SNP was seen in a single affected draft horse and the control horse. Since it was not seen in all affected draft horses, this SNP is not associated with the CPL phenotype.