Buffalobur (Solanum rostratum Dunal) () is an invasive weed in China, identifying its pathogens is crucial for developing effective biological control measures. In this study, leaf samples from S. rostratum showing typical disease symptoms were collected in Liaoning and Jilin provinces, China. The isolated fungal pathogens were identified based on their morphological characteristics and by using molecular biology techniques. Pathogenicity was assessed by artificially inoculating spore suspensions from the fungal pathogen onto the seeds, isolated leaves, and plants of S. rostratum. The safety of the fungal pathogens for eight plant species was also evaluated. We then identified the following five fungal pathogens causing disease in S. rostratum in Liaoning and Jilin provinces: Alternaria alternata, Epicoccum sorghinum, Fusarium equiseti, Curvularia hawaiiensis, and Nigrospora oryzae. These fungal pathogens exhibited pathogenicity, with N. oryzae exhibiting the strongest pathogenicity and highest safety. N. oryzae demonstrated the highest inhibition rate against the radicle germination length of S. rostratum and showed robust pathogenicity towards both isolated leaves and plants. Notably, despite inducing mild reactions in corn, sorghum, rice, and tomatoes, N. oryzae did not impose any detrimental effects on the growth of these plants.

During investigation of common linnet (Linaria cannabina) blood using the buffy coat method one bird with microfilariae in the blood was found. The morphometric description of adult worms corresponded to the Chandlerella sinensis. This species was found for the first time in common linnets. DNA sequences of cox1 and 28S gene fragments of adult worm recovered during necropsy was identical to that from the microfilariae in the bird blood. Phylogenetic analysis of the cox1 gene fragment clustered this parasite with Chandlerella quiscali. Histological examination revealed the presence of microfilariae in the lumen of small capillaries and other blood vessels in different organs, but no inflammations were notice. The greatest number of microfilariae was in the lungs. Even if there was no inflammation, but vessels associated with the lungs were markedly distended with blood, parabronchial walls were thickened and, in some cases, almost completely obstructing the lumen. The large number of microfilariae in lungs indicates possible disturbance of gas exchange in the lungs adversely affected the ability of the bird to exercise and made breathing difficult at rest. The investigation of circadian rhythm of the microfilariae showed that C. sinensis microfilariae in blood of common linnet were more numerous at night and morning and less numerous at midday. The survival rate of mosquitoes infected with C. sinensis microfilariae was significantly lower than that of uninfected mosquitoes.

Metastrongyle lungworms could be particularly detrimental for diving animals such as marine mammals; however, little is known of the drivers of pathogenic host–parasite relationships in this group. This systematic review analysed the diversity of metastrongyles in marine mammals and the host and parasite traits associated with virulence. There have been at least 40 species of metastrongyles described in 66 species of marine mammals. After penalization for study biases, Halocercus hyperoodoni, Otostrongylus circumlitus, Parafilaroides gymnurus, Halocercus brasiliensis and Stenurus minor were the metastrongyles with the widest host range. Most studies (80.12%, n = 133/166) reported that metastrongyles caused bronchopneumonia, while in the cardiovascular system metastrongyles caused vasculitis in nearly half of the studies (45.45%, n = 5/11) that assessed these tissues. Metastrongyles were associated with otitis in 23.08% (n = 6/26) of the studies. Metastrongyle infection was considered a potential contributory to mortality in 44.78% (n = 90/201) of the studies while 10.45% (n = 21/201) of these studies considered metastrongyles the main cause of death. Metastrongyle species with a wider host range were more likely to induce pathogenic effects. Metastrongyles can cause significant tissue damage and mortality in marine mammals although virulent host–parasite relationships are dominated by a few metastrongyle species with wider host ranges.

In a recent survey of nematodes associated with tobacco in Shandong, China, the root-lesion nematode Pratylenchus coffeae was identified using a combination of morphology and molecular techniques. This nematode species is a serious parasite that damages a variety of plant species. The model plant benthi, Nicotiana benthamiana, is frequently used to study plant-disease interactions. However, it is not known whether this plant species is a host of P. coffeae. The objectives of this study were to evaluate the parasitism and pathogenicity of five populations of the root-lesion nematode P. coffeae on N. benthamiana. N. benthamiana seedlings with the same growth status were chosen and inoculated with 1,000 nematodes per pot. At 60 days after inoculation, the reproductive factors (Rf = final population densities (Pf)/initial population densities (Pi)) for P. coffeae in the rhizosphere of N. benthamiana were all more than 1, suggesting that N. benthamiana was a good host plant for P. coffeae. Nicotiana. benthamiana infected by P. coffeae showed weak growth, decreased tillering, high root reduction, and noticeable brown spots on the roots. Thus, we determined that the model plant N. benthamiana can be used to study plant-P. coffeae interactions.

The world is experiencing a global push toward smart agriculture to help feed the burgeoning population by increasing food security while reducing the carbon footprint of food production. The guidelines for healthy eating have increased globally from five to seven servings of vegetables a day and this had led to the quest for a sustainable form of vegetable production that will reduce the carbon footprint and still provide consumers with the required nutrients. Microgreens contain more nutrients than some mature vegetables and can be cultivated on vertical farms, offering a different approach with the potential to resolve environmental and health challenges. Microgreens are young plantlets grown from the seeds of edible leafy vegetables and are usually eaten raw. They contain high levels of bioactive compounds and can be processed into oils to create valuable cosmetic products. Microgreens have become well-known to chefs and are gaining popularity in upmarket grocery outlets. Consequently, growing microgreens are presenting huge market opportunities worldwide. Their nutritional benefits, easy production methods and short production cycle are some of the reasons they are attractive to growers. The most important factors affecting the growth of microgreens are micro and macro-climates. One challenge to producing microgreens is that the growing environment is ideal for microbial organisms to thrive. As such, microgreens are prone to foodborne pathogens such as E. coli, Listeria and Salmonella. Consequently, the microgreens industry is facing various setbacks including product recalls from Salmonella and Listeria food poisoning outbreaks. In addition, the short shelf-life of microgreens is a serious challenge for getting microgreens to market, this is driving studies in several post-harvest treatments. This review examines the nutrient content and health benefits of microgreens and factors affecting microgreens' growth: temperature, humidity, photoperiod, fertilization, etc. and post-harvest treatments, all of which can potentially impact microbial growth, the phytochemical content and the physical appearance of microgreens bound for the market.

The success of the mollusc-parasitic nematode, Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae), as a biological control agent in Europe has led to worldwide interest in phasmarhabditids as biocontrol agents. In this study, the mass culture potential of three phasmarhabditids, namely Phasmarhabditis papillosa, Phasmarhabditis kenyaensis and Phasmarhabditis bohemica, was assessed. In addition, ten bacterial candidates, consisting of seven associated with slugs and three associated with entomopathogenic nematodes, were investigated. The bacteria were tested for their ability to cause mortality to Deroceras invadens, as well as to support nematode growth. Initial mortality studies demonstrated that Kluyvera, Aeromonas and Pseudomonas spp. (AP3) caused 100% mortality when they were injected into the haemocoel of D. invadens. However, in growth studies, Pseudomonas sp. (AP4) was found to be the most successful bacterium, leading to recovery and reproduction in almost all nematode species, except for P. kenyaensis. In flask studies, P. bohemica, which showed exceptional growth with Pseudomonas sp. (AP1), was chosen for further investigation. The effect of inoculating flasks with different concentrations of Pseudomonas sp. (AP1), as well as with different concentrations of P. bohemica, was evaluated by assessing the nematode populations for 14 days. The results indicated that the lowest, 1% (v/v), bacteria inoculation led to higher total nematode and to infective juvenile (IJ) yield, with flasks with the highest IJ inoculum (3000 IJs/ml) having a positive effect on the total number of nematodes and IJs in cultures of P. bohemica. This study presents improvements for the mass-culturing of nematodes associated with molluscs.

Root-lesion nematodes (Pratylenchus spp.) are a group of economically important pathogens that have caused serious economic losses in many crops. In 2019, root-lesion nematodes were recovered from tomato (Solanum lycopersicum) root samples collected from Sichuan Province, People's Republic of China (PRC). Extracted nematodes were disinfected, and one individual female was cultured on a carrot disc for propagation at 25 °C by parthenogenesis and designated the SC isolate. Afterwards, the isolate was identified on the basis of morphometric and molecular markers. Both morphometric characters and molecular analysis of the internal transcribed spacer region gene (ITS) of ribosomal DNA, the D2-D3 expansion region of the 28S rDNA gene and the mitochondrial cytochrome oxidase I (mtDNA-COI) gene revealed that the species of root-lesion nematode was Pratylenchus scribneri. The Bayesian tree inferred from the ITS rDNA, 28S rDNA and mtDNA-COI gene sequences also showed that this isolate formed a highly supported clade with other P. scribneri isolates. The pathogenicity of the root-lesion nematode SC isolate on tomato was assessed, showing that tomato was a suitable host for P. scribneri. To the best of our knowledge, this is the first report of P. scribneri on tomato in Sichuan Province, PRC. These are also the first molecular data obtained from P. scribneri on tomato in the PRC, and the pathogenicity of P. scribneri to tomato was studied for the first time. This study provides scientific data for the detection, identification and control of tomato root-lesion nematode disease.

Fascioliasis is a zoonotic disease caused by liver flukes transmitted by freshwater lymnaeid snails. Donkey and horse reservoir roles have been highlighted in human endemic areas. Liver fluke infection in mules has received very limited research. Their role in disease transmission, epidemiological importance and Fasciola hepatica pathogenicity are studied for the first time. Prevalence was 39.5% in 81 mules from Aconcagua, and 24.4% in 127 from Uspallata, in high-altitude areas of Mendoza province, Argentina. A mean amount of 101,242 eggs/mule/day is estimated. Lymnaeids from Uspallata proved to belong to ribosomal DNA internal transcribed spacer (ITS) markers ITS-1 and ITS-2 combined haplotype 3C of Galba truncatula. These lymnaeids were experimentally susceptible to infection by egg miracidia from mules. Infectivity, number of cercariae/snail and shedding period fit the enhanced F. hepatica/G. truncatula transmission pattern at very high altitude. This indicates that the mule is able to maintain the F. hepatica cycle independently. Individual burdens of 20 and 97 flukes were found. Mule infection susceptibility is intermediate between donkey and horse, although closer to the latter. Anatomo-pathology and histopathology indicate that massive infection may cause mule death. Haematological value decreases of red blood cells, haemoglobin, leucocytes and lymphocytes indicate anaemia and strong immunosuppression. Strongly increased biochemical marker values indicate liver function alterations. The mule probably played a role in the past exchanges with Chile and Bolivia through Mendoza province. Evidence suggests that mules could contribute to the spread of both F. hepatica and G. truncatula to human fascioliasis-endemic areas in these countries.

Staphylococci have been isolated from various sites of the body of healthy sheep, as well as from many infections of those animals, the main one being mastitis. The objective of this review is to appraise the importance and significance of staphylococci in causing mastitis in ewes. The review includes a brief classification and taxonomy of staphylococci and describes the procedures for their isolation and identification, as well as their virulence determinants and the mechanisms of resistance to antibacterial agents. Various staphylococcal species have been implicated in staphylococcal mastitis and the characteristics of isolates are discussed with regards to potential virulence factors. Staphylococcal mastitis is explicitly described, with reference to sources of infection, the course of the disease and the relevant control measures. Finally, the potential significance of staphylococci present in ewes’ milk for public health is discussed briefly.

Anastrepha ludens (Loew) (Diptera: Tephritidae) is an important pest of mango and citrus in Mexico. Here, we report the effect of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin strain MAAP1 on mortality and food consumption in A. ludens adults. Experiments were carried out under laboratory conditions at 26 ± 2 °C, 60 ± 5% RH and 12:12 photoperiod. To assess pathogenicity and sublethal effects, we sprayed flies with 1 × 109 conidia/ml and 1 × 106 conidia/ml solutions, respectively. Food consumption was measured by providing known amounts of food through capillary tubes to caged flies. We found that M. anisopliae was highly virulent to A. ludens, causing 92.3 ± 0.62% mortality at day 13 post-exposure. Interestingly, food consumption reduced by 17% in flies exposed to the fungus. These results support the implementation of M. anisopliae within current strategies used for the integrated pest management of A. ludens.

The amphibian pathogen Batrachochytrium dendrobatidis (Bd) has recently emerged as a primary factor behind declining global amphibian populations. Much about the basic biology of the pathogen is unknown, however, such as its true ecological niche and life cycle. Here we evaluated invertebrates as infection models by inoculating host species that had previously been suggested to be parasitized in laboratory settings: crayfish (Procambarus alleni) and nematodes (Caenorhabditis elegans). We found neither negative effects on either host nor evidence of persistent infection despite using higher inoculum loads and more pathogen genotypes than tested in previous studies. In contrast, addition of Bd to C. elegans cultures had a slight positive effect on host growth. Bd DNA was detected on the carapace of 2/34 crayfish 7 weeks post-inoculation, suggesting some means of persistence in the mesocosm. These results question the role of invertebrates as alternative hosts of Bd and their ability to modulate disease dynamics.

Amoebae were isolated from contact lenses of a symptomatic lens wearer in Spain. Protozoa were characterized by studying their morphology, biology, protease activity and the 18S rRNA gene sequence. Morphology of the organism was observed by light microscopy, scanning electron microscopy and transmission electron microscopy. Its structure corresponded to an amphizoic amoeba. The protozoa grew well at 37 °C and poorly at lower temperatures. In addition, it was capable of lysing mammalian cells in vitro. A major 56 kDa proteolytic enzyme was observed in amoeba crude extracts by gelatin–sodium dodecyl sulphate–polyacrylamide gel electrophoresis. Most proteolytic enzymes in protozoa extracts showed significant activity over a wide range of pH (3–9) and temperature (8–45 °C) values. The assays on inhibition of protease activity indicated strongly that enzymes detected in amoeba extracts corresponded to serine proteases and, to a lesser extent, cysteine proteases. The use of proteinase inhibitors on a tissue culture model proved that the proteinase activity is critical for developing focal lesions in HeLa cell monolayers. Finally, partial sequencing of the 18S ribosomal RNA gene and phylogenetic analyses indicated that the isolate is closely related to Acanthamoeba griffini H37 from the UK (T3 genotype).

Members of the family Trichomonadidae, mainly Trichomonas gallinae and Tetratrichomonas gallinarum, represent important parasites in birds with worldwide presence, since being reported in the 19th century. Especially Columbiformes, Falconiformes and Strigiformes can be severely affected by trichomonads, whereas the majority of infections in Galliformes and Anatiformes are subclinical although severe infections are occasionally reported. With the recent appearance of deadly infections in wild Passeriformes the protozoan parasite T. gallinae obtained greater attention which will be addressed in this review. Although light microscopy remains the method of choice to confirm the presence of trichomonads molecular studies were introduced in recent years, in order to characterize the parasites and to establish relationships between isolates. Isolation of trichomonads is a prerequisite for detailed in vitro and in vivo studies and different media are reported to obtain suitable material. The limited information about virulence factors will be reviewed in context with the pathogenicity of trichomonads which varies greatly, indicating certain strain heterogeneity of the parasites. Options for treatment characterized by the leading role of imidazoles whose activity is sometimes hampered by resistant parasites remains a challenge for the future. Introducing more standardized genetic studies and investigations concentrating on the host-pathogen interaction should be helpful to elucidate virulence factors which might lead to new concepts of treatment.

The necessity to easily establish Histomonas meleagridis cultures has been underlined extensively by many researchers in order to gain more insights in the biology of H. meleagridis. In addition the occurrence of different protozoa in the caeca of birds impedes, however, the isolation and propagation of H. meleagridis from field outbreaks. Therefore, in a kinetic study using transmission electron microscopy the deleterious effects of adventitious protozoa including Tetratrichomonas gallinarum and Blastocystis spp. on cultured H. meleagridis were examined. To overcome this issue, an easy and successful approach to establish the mono-eukaryotic H. meleagridis culture free of other host's protozoa is proposed. At 10 days post infection, liver lesions of H. meleagridis-infected birds were isolated and inoculated into culture media pre-incubated with caecal bacteria. After 48 h of incubation, presence of H. meleagridis in the cultures was confirmed through morphological evaluation. Additionally, TEM examination and analysis by PCR amplification of the small subunit rRNA gene could exclude the co-cultivation of T. gallinarum and Blastocystis spp. Furthermore, after successful propagation and maintenance of the cultured H. meleagridis, its pathogenicity was affirmed in an infection experiment in turkeys.

Canine influenza virus (CIV) is an emerging pathogen that causes acute respiratory disease in dogs. The aim of this study was to investigate the pathogenicity of A/canine/Jiangsu/06/2010 (H3N2) virus isolated in China. Nine dogs were inoculated intranasally with 107·95 of 50% egg infectious dose (EID50) of the virus. The onset of clinical signs and virus shedding was observed on day 1 post-infection (p.i.). The peak clinical score occurred between days 4 and 6 p.i. The experimentally infected dogs were found to shed virus not only via the respiratory tract but also via the digestive tract. Viral RNA could be detected in multiple organs including the trachea, lung, liver, spleen, kidney, brain and duodenum. All the sampled organs from infected dogs showed significant lesions and viral antigen staining. The results differed from those reporting using previous CIV strains; the Chinese isolate could induce extrapulmonary infection and cause extensive lesions in dogs.

Regulations in the United States require animal biosafety level 3 (ABSL-3) or biosafety level 3 agriculture (BSL-3-Ag) containment for many endemic zoonotic pathogens and etiologic agents of foreign animal diseases. In an effort to protect public health, billions of dollars were invested in regulatory programs over many years to reduce the prevalence of zoonotic pathogens such as Brucella and Mycobacterium bovis in domestic livestock. In addition to research needs in domestic livestock hosts, the establishment of brucellosis and tuberculosis in wildlife in the United States has created a need for research studies addressing these zoonotic diseases. As guidelines in the Biosafety in Microbiological and Biomedical Laboratories (BMBL, 2009) for BSL-3 and BSL-3-Ag facilities are primarily directed toward laboratory or vivarium facilities, additional issues should be considered in designing large animal containment facilities for domestic livestock and/or wildlife. Flight distance, herd orientation, social needs, aggressiveness, and predictability are all factors we considered on a species by species basis for designing our containment facilities and for work practices with large ruminants. Although safety risk cannot be completely eliminated when working with large animals, studies in natural hosts are critical for advancing vaccine and diagnostic development, and providing basic knowledge of disease pathogenesis in natural hosts. Data gathered in these types of studies are vital for state and national regulatory personnel in their efforts to design strategies to control or eradicate diseases such as brucellosis and tuberculosis in their natural hosts, whether it is domestic livestock or wildlife. It is likely that failure to address the prevalence of disease in wildlife reservoirs will lead to re-emergence in domestic livestock. The overall benefit of these studies is to protect public health, provide economic benefits to producers, and protect the economic investment made in regulatory programs.

Humans and other hosts acquire Toxoplasma gondii infection by ingesting tissue cysts in undercooked meat, or by food or drink contaminated with oocysts. Currently, there is no vaccine to prevent clinical disease due this parasite in humans, although, various T. gondii vaccine candidates are being developed. Mice are generally used to test the protective efficacy of vaccines because they are susceptible, reagents are available to measure immune parameters in mice, and they are easily managed in the laboratory. In the present study, pathogenesis of toxoplasmosis was studied in mice of different strains, including Human Leukocyte Antigen (HLA) transgenic mice infected with different doses of T. gondii strains of different genotypes derived from several countries. Based on many experiments, the decreasing order of infectivity and pathogenicity of oocysts was: C57BL/6 background interferon gamma gene knock out (KO), HLA-A*1101, HLA-A*0201, HLA-B*0702, Swiss Webster, C57/black, and BALB/c. Mice fed as few as 1 oocyst of Type I and several atypical strains died of acute toxoplasmosis within 21 days p.i. Some Type II, and III strains were less virulent. The model developed herein should prove to be extremely useful for testing vaccines because it is possible to accurately quantitate a challenge inoculum, test the response to different strains of T. gondii using the same preparations of oocysts which are stable for up to a year, and to have highly reproducible responses to the infection.

Whole blood collected from koalas admitted to the Australian Zoo Wildlife Hospital (AZWH), Beerwah, QLd, Australia, during late 2006–2009 was tested using trypanosome species-specific 18S rDNA PCRs designed to amplify DNA from Trypanosoma irwini, T. gilletti and T. copemani. Clinical records for each koala sampled were reviewed and age, sex, blood packed cell volume (PCV), body condition, signs of illness, blood loss, trauma, chlamydiosis, bone marrow disease, koala AIDS and hospital admission outcome (‘survival’ / ‘non-survival’) were correlated with PCR results. Overall 73·8% (439/595) of the koalas were infected with at least 1 species of trypanosome. Trypanosoma irwini was detected in 423/595 (71·1%), T. gilletti in 128/595 (21·5%) and T. copemani in 26/595 (4·4%) of koalas. Mixed infections were detected in 125/595 (21%) with co-infections of T. irwini and T. gilletti (101/595, 17%) being most common. There was a statistical association between infection with T. gilletti with lower PCV values and body condition scores in koalas with signs of chlamydiosis, bone marrow disease or koala AIDS. No association between T. gilletti infection and any indicator of health was observed in koalas without signs of concurrent disease. This raises the possibility that T. gilletti may be potentiating other disease syndromes affecting koalas.

Infectivity of the entomopathogenic fungus Metarhizium anisopliae against workers of the arboreal termite Odontotermes sp. was assayed under laboratory conditions. Test isolates were collected from different sources, including soil from varied locations and insect hosts from the orders Lepidoptera, Coleoptera, Hemiptera and Orthoptera. All the 23 isolates tested and the standard (ARSEF 7413) were pathogenic to the workers of Odontotermes sp. at a concentration of 107 conidia/ml, with mean mortality ranging from 57.5 to 100%. Two of the isolates (Ma2, Ma13) and the standard caused 100% mortality in the termite species. A detailed bioassay was subsequently conducted with the five most promising isolates, namely Ma1, Ma2, Ma13, Ma16 and Ma17, at concentrations ranging from 104 to 107 conidia/ml. The lethal concentrations (LC50) of these isolates ranged from 0.01 to 0.46 × 105 conidia/ml. The average survival time (AST) for the termites treated with the most virulent isolate (Ma2) varied from 4.2 to 5.7 days across the four spore loads, while AST with the standard isolate ranged from 5.3 to 6.3 days. Two of the isolates, Ma2 and Ma13, were found to be significantly more pathogenic to Odontotermes sp. workers than all the others, including the standard.

Male Hisex chicks were used in two experiments to investigate the interaction between Ascaridia galli infection and supplemental copper from basic and neutral salts. This was assessed by means of body weights, mortality, parasite burden and liver copper level. Cu2(OH)3Cl reduced the number of parasites but CuSO4.5H2O and CuCO3.Cu(OH)2.nH2O did not affect the parasite burden.