Experimental challenge models for Johne's disease: a review and proposed international guidelines

An international committee of Johne's disease (JD) researchers was convened to develop guidelines for JD challenge studies in multiple animal species. The intent was to develop and propose international standard guidelines for models based on animal species that would gain acceptance worldwide. Parameters essential for the development of long-term and short-term infection models were outlined and harmonized to provide a "best fit" JD challenge model for cattle, goats, sheep, cervids, and mice. These models will be useful to study host-pathogen interactions, host immunity at the local and systemic level, and for evaluating vaccine candidates and therapeutics. The consensus guidelines herein list by animal species strains of Mycobacterium avium subsp. paratuberculosis used, challenge dose, dose frequency, age of challenge, route of challenge, preparation of inoculum, experimental animal selection, quality control, minimal experimental endpoints and other parameters.     

Forest structure and woody plant species composition after a wildfire in beech forests in the north of Iran

Beech (Fagus orientalis Lipsky) forest covers about 565,000 ha of land in Guilan province, north of Iran and forms a major carbon pool. It is an important economic, soil protection and recreation resource. We studied long-term effects of fire on the structure and composition 37 years after fire occurrence in these forests. To do this research, we selected 85 ha burned and 85 ha unburned beech forests). The results indicated that the fire had not changed the overall uneven-aged structure, but it changed forest composition from pure stands to mixed stands that now include species such as Carpinus betulus, Acer cappadocicum and Alnus subcordata. The density of trees and regeneration was significantly increased, while the density of shrubs significantly decreased. The main reasons for increased tree regeneration were attributed to (1) reduction of litter depth, and (2) increase in available light from opening of the canopy and reduction in shrub competition. It is apparent that the forest is on a path to return to its natural state before the fire after 37 years.     

Effect of temperature on the expression of cytoplasmic male sterility in cultivated barley (Hordeum vulgare L.)

Twenty‐three cytoplasmic male sterile BC₁F₁ barley lines were exposed to varying temperature treatments (TTs) to examine the effect of temperature on the stability of the expression of cytoplasmic male sterility (cms). The TTs used for this test were: (i) controlled low‐temperature treatment of 16°C (CL), (ii) controlled medium‐temperature treatment of 21–24/16–17°C day/night (CM) and (iii) ambient glasshouse‐temperature treatment of 24–41/16–17°C day/night (AG). The expression of cms was found to be variably influenced by temperature and by the genetic background of the cms recipient lines. Ten cms lines exhibited consistently complete male sterility over TTs, indicating that these lines are completely under cms genes control, while other lines showed partial revision to fertility across different TTs with profound influence of warm temperatures (CM and AG) in breaking down the cms in barley.     

Role of sponge gourd in apical leaf curl disease of potato in Northern India

Potato (Solanum tuberosum) is one of the important vegetable crops in the world and its production is seriously affected by apical leaf curl disease in northern India. This paper reveals the role of cucurbits in maintaining Tomato leaf curl New Delhi virus (ToLCNDV) and Potato apical leaf curl (PALCD) disease in that region. The affected plants showed severe leaf curling and stunted growth. The begomovirus causing leaf curling and mosaic disease in cucurbits could be easily transmitted by the whitefly to potato crops and develop apical leaf curl disease in northern India. The movement of the virus by whiteflies from cucurbits to potato and tomato is possible because of overlapping of planting and harvesting dates of these crops. The causal virus was identified as a begomovirus on the basis of whitefly transmission, PCR, dot blot hybridization, cloning and sequencing of the coat protein gene. The comparison of full length coat protein gene sequence homology revealed that 90% identity with the coat protein gene of ToLCNDV- [Luffa] isolate and the phylogenetic tree derived from these sequences with other selected begomoviruses formed a close cluster with ToLCNDV isolates. The findings proved that the virus causing disease in cucurbits could easily move to tomato and potato and cause leaf curl disease naturally. This is the first observation on the role of sponge gourd for maintenance of ToLCNDV and serving as a host for PALCD in northern India. The findings indicate that the causal organism is a strain of ToLCNDV.     

Brucellosis in camels

Camels are highly susceptible to brucellosis caused by Brucella melitensis and Brucella abortus. Difficulties can arise in diagnosis of camel brucellosis, especially as this disease provokes only few clinical signs in contrast to its clinical course in cattle. Because none of the commonly used serological test can be perceived as a perfect test for Brucella diagnosis in camel and most serological tests used for camels have been directly transposed from cattle without adequate validation, an incorrect diagnosis may occur when diagnosis is based on serology alone. Of imminent concern is the fact that brucellosis can be easily transmitted from animals or their products to humans mainly via milk. In many developing countries in the arid areas of Asia and Africa, camels are still the most important productive livestock for nomadic populations. Therefore, we reviewed the literatures on camel brucellosis to highlight the epidemiologic, economic and public health impact of camel brucellosis as a basis for designing effective control strategies.     

Evaluation of different diagnostic methods for diagnosis of Lumpy skin disease in cows

Viral isolation, polymerase chain reaction (PCR), dot blot hybridization (DBH), and indirect enzyme-linked immunosorbent assay (iELISA) were used for the diagnosis of lumpy skin disease in clinically infected, fevered, and apparently normal dairy cows. Lumpy skin disease virus (LSDV) was isolated from skin biopsies and blood samples collected from clinically infected cows in percentages of 72% and 20%, respectively. The virus recovered from blood samples collected from fevered cows in percentage of 33.3%. Both PCR and DBH detected viral DNA in 100% of skin biopsies collected from clinically infected cows whereas the detection rates in blood samples collected from clinically infected animals were 100% and 84% using PCR and DBH, respectively. Viral DNA was detected in blood samples collected from fevered cows using PCR and DBH in percentages of 77.8% and 66.6%, respectively. Only 19.1% of blood samples collected from in-contact cows was positive for both of PCR and DBH. Detection rates of antibodies against LSDV using iELISA in serum samples collected from clinically infected and fevered cows were 56% and 11.1%, respectively, whereas all in-contact cows had no antibodies against the virus.     

Different Morphologies of Leishmania major Amastigotes with No Molecular Diversity in a Neglected Endemic Area of Zoonotic Cutaneous Leishmaniasis in Iran

Background:

Molecular diversity of Leishmania major and its morphological changes have become a controversial issue among researchers. Some aspects of polymorphic shapes of amastigotes in clinical manifestations along with molecular variation were evaluated among suspected patients of some exceptional zoonotic cutaneous leishmaniasis locations in Northern Khuzestan, Southwestern Iran.

Methods:

Suspected patients (n = 165) were sampled in zoonotic cutaneous leishmaniasis foci over two consecutive years during 2012-2014. Prepared smears were stained, scaled and measured by ocular micrometer. DNA was extracted from smears; ITS-rDNA and Cytochrome b (Cyt b) markers were amplified, and PCR products were digested by BsuR1 restriction enzyme. Then the RFLP and sequencing were employed.

Results:

Only L. major was identified in patients containing regular amastigotes'' shapes (oval or round) with a size of 2-4 µm in each of classical wet, dry, mixed lesions. Meanwhile, irregular shapes (spindle, pear, or cigarette) were observed separately in non-classical wet lesions with more than 4 µm. Interestingly, a few amastigotes with an external flagellum were observed in some lesions. All sequenced ITS-rDNA and Cyt b genes of L. major did not show any molecular variation (χ ² P > 0.05), including only one common haplotype (GenBank access no. {"type":"entrez-nucleotide","attrs":{"text":"EF413075","term_id":"151936076","term_text":"EF413075"}}EF413075).

Conclusion:

Findings proved that unlike other endemic foci, there is not a meaningful correlation between phenotypic and genotypic features of L. major isolates. This study is considered as the first comprehensive report to incriminate morphometric shapes of L. major amastigotes, which enhances our knowledge concerning their relevance with various clinical appearances and genotypic traits. Key Words: Leishmania major, Nuclear gene, Mitochondrial gene, Amastigote shapes, Iran