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The stable fly, Stomoxys calcitrans L. (Diptera: Muscidae), is a significant insect pest with global veterinary implications due to its capacity to both cause nuisance and transmit disease-causing pathogens to livestock. This study aimed to determine the livestock bedding preferred for use as a development substrate by S. calcitrans larvae and field-collected adults. The result showed that S. calcitrans larvae exhibited a preference (26.7%) for 7-day-old cow manure. Gravid females displayed a pronounced preference (55.0%) for fresh cow manure. As there were eight choices, indifference would result in 12.5% for each bedding substrate. Furthermore, the efficacy of four entomopathogenic nematodes (EPNs), namely Heterorhabditis bacteriophora (Poinar), Heterorhabditis indica Poinar, Karunakar & David (Rhabditida: Heterorhabditidae), Steinernema siamkayai Poinar, Karunakar & David and Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae), against S. calcitrans larvae and the persistence after application to livestock bedding substrates were evaluated under laboratory conditions. In filter paper bioassays, all four EPNs caused 76.7%-100.0% mortality in the second instar larvae of S. calcitrans when applied at 50 and 100 infective juveniles (IJs)/cm 2 within 5 days after exposure. For the third instar larvae of S. calcitrans, only H. indica induced high mortalities of 86.6% when applied at 100 IJs/cm 2 within 5 days after exposure, while the other EPNs resulted in mortalities of less than 70%. The data further demonstrated that H. bacteriophora, H. indica and S. siamkayai remained present in the substrates linked to S. calcitrans larvae for as long as 7 days after the application of EPNs. This study demonstrates the potential of EPNs as a biologically based control agent against larvae of S. calcitrans, a serious pest and significant vector for various livestock animals., (© 2024 Royal Entomological Society.)

Northeastern Thailand relies on agriculture as a major economic activity, and has used high levels of agrochemicals due to low facility, and salty sandy soil. To support soil recovery and sustainable agriculture, local farmers have used organic fertilizers from farmed animal feces. However, knowledge about these animal fecal manures remains minimal restricting their optimal use. Specifically, while bacteria are important for soil and plant growth, an abundance and a diversity of bacterial composition in these animal fecal manures have not been reported to allow selection and adjustment for a more effective organic fertilizer. This study thereby utilized metagenomics combined with 16S rRNA gene quantitative PCR (qPCR) and sequencing to analyze quantitative microbiota profiles in association with nutrients (N, P, K), organic matters, and the other physiochemical properties, of the commonly used earthworm manure and other manures from livestock animals (including breed and feeding diet variations) in the region. Unlike the other manures, the earthworm manure demonstrated more favorable nutrient profiles and physiochemical properties for forming fertile soil. Despite low total microbial biomass, the microbiota were enriched with maximal OTUs and Chao richness, and no plant pathogenic bacteria were found based on the VFDB database. The microbial metabolic potentials supported functions to promote crop growth, such as C, N and P cyclings, xenobiotic degradation, and synthesis of bioactive compounds. Pearson's correlation analyses indicated that the quantitative microbiota of the earthworm manure were clustered in the same direction as N, and conductivity, salinity, and water content were essential to control the microbiota of animal manures.

The loss of biodiversity-caused mainly by habitat destruction-is one of the environmental problems with major repercussions on ecosystem functioning. Nevertheless, our understanding of the functional consequences of habitat changes on the communities and ecosystems remains limited to a small number of case studies. We evaluated the change in taxonomic and functional diversity of copro-necrophagous beetles (Scarabaeinae) and their relationship with the varying environmental factors present in four habitats with different degrees of disturbance. Furthermore, we evaluated how changes in taxonomic and functional diversity affect the rates of excrement removal. The collections were carried out at four locations in the state of Santa Catarina, Southern Brazil, on natural systems with different degrees of disturbances (forests in advanced and initial succession) and agroecosystems (silviculture and pastures dedicated to livestock). We collected a total of 1266 dung beetles distributed in 35 species and classified into 11 functional groups. The taxonomic and functional diversity analyses showed that habitats that still maintain an arboreal stratum do not present differences between them, in contrast to habitats dedicated to livestock where there was a significant loss of species and functional groups. The distance between the trees, as well as the air and soil temperatures were determining factors in the selection of species and functional groups. Some of these environmental factors explain the differences in functional traits, represented as varying abundances of the species found. The rates of manure removal from the ecosystem were positively correlated to taxonomic and functional richness as well as biomass of beetles. Thus, we can conclude that habitats with tree strata have the capacity to preserve a larger proportion of the regional set of species as well as the important ones, while preserving the taxonomic and functional diversity and the ecosystem functions, such as the excrement removal rate., Competing Interests: The authors have declared that no competing interests exist.

Organic waste is a rapidly increasing problem due to the growth of the agricultural production needed to meet global food demands. Development of sustainable waste management solutions is essential. Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) (BSF), larvae are voracious consumers of a wide range of organic materials ranging from fruits and vegetables to animal remains, and manure. Thanks to this ability and considering the larval high protein and lipid content, BSF larvae are a useful additive in animal feeds and biodiesel production. Unfortunately, the feasibility of using the black soldier fly as a tool for waste valorization and feed production has primarily been investigated at the benchtop scale. Thus, mobilization of current practices to an industrial scale is challenging because scaling up from small laboratory studies to large industrial studies is not necessarily linear. The goal of this study was to demonstrate the ability of the BSF to recycle organic waste at an industrial scale. To accomplish this goal, three organic waste streams were used (e.g., apples, bananas, and spent grain from a brewery) to test six diet treatments (1) apple, (2) banana, (3) spent grain, (4) apple and banana, (5) apple and spent grain, and (6) banana and spent grain. Working at scale of 10,000 BSF larvae life history traits, waste valorization, protein and lipid profiles were measured for each diet treatment. Differences were recorded across all variables, except substrate conversion, for larvae fed on fruit and spent grain (alone or with fruit). Growth rate significantly differed across treatments; larvae reared on spent grain grew twice as fast as those fed apples alone, but those reared on the apple and spent grain mixture produced twice as much insect biomass. However, it should be noted that larvae resulting from the apple diet contained 50% more fat than larvae fed the fruit and spent grain mixtures. Commonly-available organic wastes were successfully used at an industrial scale to produce BSF larvae that have the potential to substitute other sources of protein and lipids in different industrial applications. Industrialization efforts are encouraged to assess these impacts when integrating diverse ingredients into larval diets as a means to more precisely predict output, such as larval development time and final larval biomass.

Antibiotics are commonly used in livestock and poultry farming. Residual antibiotics in manure may lead to antibiotic pollution of soil, surface water, and groundwater through land application and run-off rainfall. The black soldier fly (BSF) Hermetia illucens is a good candidate for manure management. We evaluated the effect of sulfonamide pollution on the growth of H. illucens. Four treatments were considered with a sulfonamide content in the feed of 0 (control group), 0.1, 1, and 10 mg/kg. The control larvae were fed without sulfonamide. Survival and development status of the individuals were recorded daily. The weights of the fifth instar larvae, prepupae, and pupae were checked. Antioxidant enzyme activity was determined with the fifth instar larvae. The results showed that a low (0.1 and 1 mg/kg) concentration of sulfonamides had no effects on larval survival, pupation, and eclosion of BSFs. A high sulfonamide concentration of 10 mg/kg had a significant effect on the survival of larvae and pupae and on the body weight of larvae, prepupae and pupae. Peak of the cumulated pupation rate and eclosion rate in the sulfonamide treatment of 10 mg/kg was very low. Pupation and eclosion in this group peaked later than that of the control and low sulfonamide concentration treatment groups (0.1 mg/kg and 1 mg/kg). Larvae from the sulfonamides group showed lower antioxidase activities than that of the control. In sulfonamide groups, the activity of peroxidase and superoxide dismutase was reduced in a concentration-dependent manner. Sulfamonomethoxine, sulfamethoxazole, and sulfamethazine were not detected in the harvested prepupae. Only sulfadiazine was discovered in the sulfonamide treatments of 1 and 10 mg/kg. In conclusion, BSFs can tolerate certain concentrations of sulfonamide contamination., Competing Interests: The authors have declared that no competing interests exist.

Spent poultry litter use as a fertilizer in horticulture supports stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) development. Stable fly continues to have an economic impact on livestock production and rural lifestyle in south-western Australia. The use of raw poultry manure is banned in 12 Shires surrounding Perth. The loss of market options for West Australian broiler growers has caused economic hardship. Hence, this study examined a range of chemical and biological amendments to spent poultry broiler litter in preventing stable fly and nuisance fly development. These included alkalizers (i.e., lime sand, quicklime, soda ash, and shell grit), acidifiers (aluminum sulfate, sodium bisulfate), gypsum, zeolite, spongolite, calcium cyanamide, and two fungal agents. The treated litters were placed under irrigation in horticulture with amendments added prior to them being exposed in the field as replicate 1-liter pads. In total, 19,559 stable flies developed from the spent litters exposed over five field experiments (88.7% of all flies recovered). House flies (Musca domestica L. (Diptera: Muscidae); 2,067 or 9.4%), false stable flies (Muscina stabulans Fallén (Diptera: Muscidae); 414 or 1.9%), and two sarcophagids (flesh fly) also developed from the litter. Borax completely prevented any fly development from the litter. Calcium cyanamide (1-2.5% v/v) and sodium bisulfate (10%) reduced stable fly numbers by as much as 99-100% when added to litter. Alkalizers, zeolite, spongolite, and entomopathogenic fungi had no significant impact on stable fly development. The addition of either calcium cyanamide or sodium bisulfate to raw litter can boost the fertilizer value of the litter while preventing stable fly development.

Houseflies (Musca domestica) spend part of their life development on animal or human manure. Manure is high in pathogenic microbes; thus, houseflies have been known as a mechanical vector for various important zoonotic diseases. Therefore, the present study showcases captured houseflies from intensive swine production regions (which are areas of high manure concentration) in Southern Brazil, and analyses their bodies' to the presence of Escherichia coli and Salmonella sp. and the sensitivity of these bacteria to various antibiotics. Additionally, Quantitative Microbiology Risk Assessment was performed simulating the contamination of lettuce by flies' bacteria and subsequent lettuce consumption by an adult human being. Houseflies were captured in swine buildings and farm houses from five farms. E. coli quantification values ranged from 10 4 to 10 6 CFU/20 flies, and all sampling sites had positive results from bacteria presence in the collected houseflies. On the other hand, Salmonella sp. presence was observed in only three farms, where the quantification ranged from 10 2 to 10 5 CFU/20 flies. The bacteria showed to be resistant to at least two from the four tested antibiotics (ampicillin, Cefalotin, Ciprofloxacin and Norfloxacin) antibiotics used in human or veterinary medicine. Infection probability analyses showed risk of human infection by E.coli, indicating possible transmission of zoonotic pathogens through flies. In this context, it was possible to conclude that there is a need for flies control, especially in swine farms where zoonotic pathogens can be abundant, to minimize the health impact of the vectorization of enteric bacteria., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The study investigated the risks arising from application of pig slurry to soil in relation to viability of Salmonella typhimurium, Escherichia coli, total coliforms, faecal enterococci and eggs of Ascaris suum at different temperatures. Potential effect of changes in physico-chemical parameters, particularly dry matter (DM), pH and ammonia, were also investigated. Examination showed that S. typhimurium was devitalised after storage in the slurry for 115 days at 4 °C and after 90 days at 20 and 42 °C. Devitalization of E. coli and faecal entrerococci required more than 115 at temperature of 4 °C and faecal enterococci were recovered from slurry after 115 days of storage even at temperature of 20 °C. Total coliforms survived for 115 days at all investigated temperatures. Complete devitalization of A. suum eggs was not achieved even after 115 days at 42 °C. Our investigations indicated potential microbial and parasitic risk related to application of pig slurry to soil even after 115 days of storage.

Chrysomya megacephala is a saprophagous fly whose larvae can compost manure and yield biomass and bio-fertilizer simultaneously. However, there are concerns for the safety of the composting system, that is risk of diseases spread by way of manure pathogens, residue of harmful metals and emission of greenhouse gases. Microbiota analysis and heavy metal speciation by European Communities Bureau of Reference were evaluated in raw, C. megacephala-composted and natural stacked swine manure to survey pathogenic bacterial changes and mobility of lead and cadmium in manure after C. megacephala feeding; the emission rate of CH 4 and N 2 O from manure during C. megacephala composting and natural stacking was also measured. C. megacephala composting altered manure microbiota, reduced the risk of pathogenic bacteria and maintained the stability, and microbiota changes might be associated with heavy metal fractions, especially in Pseudomonas and Prevotella. In addition, C. megacephala-composting significantly reduced the emission rate of CH 4 and N 2 O in comparing with natural stacking situation and the first two days should be the crucial period for CH 4 and N 2 O emission measurement for manure treatment by C. megacephala. Moreover, OTU26 and Betaproteobacteria were changed after C. megacephala composting which might play a role in emission of CH 4 and N 2 O, respectively., (© 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.)

Background: Black soldier fly larvae are converters of organic waste into edible biomass, of which the composition may depend on the substrate. In this study, larvae were grown on four substrates: chicken feed, vegetable waste, biogas digestate, and restaurant waste. Samples of prepupae and substrates were freeze-dried and proximate, amino acid, fatty acid and mineral analyses were performed., Results: Protein content of prepupae varied between 399 and 431 g kg -1 dry matter (DM) among treatments. Differences in amino acid profile of prepupae were small. On the other hand, the ether extract (EE) and ash contents differed substantially. Prepupae reared on digestate were low in EE and high in ash (218 and 197 g kg -1 DM, respectively) compared to those reared on vegetable waste (371 and 96 g kg -1 DM, respectively), chicken feed (336 and 100 g kg -1 DM, respectively) and restaurant waste (386 and 27 g kg -1 DM, respectively). Prepupal fatty acid profiles were characterised by high levels of C12:0 in all treatments., Conclusion: Since protein content and quality were high and comparable for prepupae reared on different substrates, black soldier fly could be an interesting protein source for animal feeds. However, differences in EE and ash content as a function of substrate should be considered. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Records at the Endulen Hospital in the Ngorongoro Conservation Area (NCA), Tanzania, reveal that soil-transmitted helminth infections and protozoa are consistently in the top ten diagnoses for Maasai pastoralists, indicating a significant public health concern. Nevertheless, Maasai pastoralist adaptations to life in close proximity to livestock and to unreliable access to water raise important questions about experiences of, and resiliency to, parasitic infections. Though these infections are particularly prevalent among youth in low- and middle-income countries (LMIC), a focus on resiliency highlights local capacity to recover from and prevent illness. For instance, how is human parasitism perceived and experienced among communities displaying behaviours that studies have associated with transmission of diarrhoeal diseases, such as open defecation? Among these communities, how is parasitism seen to impact the health and development of children? And, what resources are available to endure or mitigate this heavy disease burden among affected communities? This study draws on formative research carried out in May 2014 in anticipation of an innovative school-based and youth-driven water, sanitation and hygiene education intervention rolled out in two boarding schools in the NCA in subsequent months. The initiative is grounded in a One Health approach to health promotion, drawing on partnerships in medicine, public health and veterinary medicine to appreciate the unique interactions between humans, animals and the environment that shape well-being among pastoralist communities. Qualitative data generated through group discussions with secondary school youth (n=60), Maasai teachers (n=6) and a women's group (n=8) in the NCA convey existing knowledge of the prevalence, prevention and treatment of human parasitism. An underlying principle of the larger initiative is to engage youth as creative agents of change in developing and sustaining locally relevant health promotion strategies. Findings highlight practical knowledge around certain 'neglected tropical diseases', namely helminths, among pastoralist communities in the NCA, in turn feeding into the development of the science fair and related interventions.

Manure application is a source of pathogens to the environment. Through overland runoff and tile drainage, zoonotic pathogens can contaminate surface water and streambed sediment and could affect both wildlife and human health. This study examined the environmental occurrence of gene markers for livestock-related bacterial, protozoan, and viral pathogens and antibiotic resistance in surface waters within the South Fork Iowa River basin before and after periods of swine manure application on agricultural land. Increased concentrations of indicator bacteria after manure application exceeding Iowa's state bacteria water quality standards suggest that swine manure contributes to diminished water quality and may pose a risk to human health. Additionally, the occurrence of HEV and numerous bacterial pathogen genes for Escherichia coli, Enterococcus spp., Salmonella sp., and Staphylococcus aureus in both manure samples and in corresponding surface water following periods of manure application suggests a potential role for swine in the spreading of zoonotic pathogens to the surrounding environment. During this study, several zoonotic pathogens were detected including Shiga-toxin producing E. coli, Campylobacter jejuni, pathogenic enterococci, and S. aureus; all of which can pose mild to serious health risks to swine, humans, and other wildlife. This research provides the foundational understanding required for future assessment of the risk to environmental health from livestock-related zoonotic pathogen exposures in this region. This information could also be important for maintaining swine herd biosecurity and protecting the health of wildlife near swine facilities., (Published by Elsevier B.V.)

Toxocara canis, Toxocara cati and Ascaris suum are worldwide-distributed zoonotic roundworms of dogs, cats and pigs, respectively. The epidemiology of these parasites in developed countries is largely unclear. Two countrywide cross-sectional serosurveys were therefore conducted in the Netherlands in 1995/1996 and 2006/2007 to investigate the prevalence, trends and risk factors for human Toxocara and Ascaris infections in the general population. The Netherlands is characterized by high pig production, freedom from stray dogs and virtual absence of autochthonous infections with the human-adapted roundworm Ascaris lumbricoides. Over the 10 years between the two serosurveys, Toxocara seroprevalence decreased significantly from 10.7 % (n = 1159) to 8.0 % (n = 3683), whereas Ascaris seroprevalence increased significantly from 30.4 % (n = 1159) to 41.6 % (n = 3675), possibly reflecting concomitant improvements in pet hygiene management and increased exposure to pig manure-contaminated soil. Increased anti-Toxocara IgGs were associated with increasing age, male gender, contact with soil, ownership of cats, cattle or pigs, hay fever, low education, high income and non-Western ethnic origin. Increased anti-Ascaris IgGs were associated with increasing age, owning pigs, low education, childhood geophagia and non-Dutch ethnic origin. Besides identifying specific groups at highest risk of Toxocara and Ascaris infections, our results suggest that these infections mainly occur through environmental, rather than foodborne, routes, with direct contact with soil or cat and pig ownership being potentially modifiable exposures.

Insects rely mainly on their well-developed and highly sophisticated olfactory system to discriminate volatile cues released from host and nonhost substances, mates, oviposition substrates, and food sources. Onset of first mating, mating duration, and onset of first oviposition, oviposition period, fecundity (number of eggs laid by a female), and longevity of freshly emerged Musca domestica L. (Diptera: Muscidae) adults were observed in the presence of different animal manures: cow, horse, donkey, poultry, and an artificial diet. The M. domestica adults exposed to horse manure showed a delay in onset of first mating and first oviposition, prolonged mating duration, and reduced fecundity compared to the artificial diet (control). Likewise, the fecundity was reduced in the presence of donkey manure as compared to artificial diet. The onset of first mating was delayed and duration of first mating was shortened in the presence of cow manure as compared to artificial diet and no oviposition was observed throughout the duration of the experiment. However, the reproductive behaviors and all fitness measures in adults exposed to poultry manure were similar or even better, compared to the artificial diet. Surprisingly, in a free-choice attraction assay, the highest numbers of adult flies were attracted toward the cow manure as compared to all other manures as well as the artificial diet. However, the numbers of flies captured in all other types of manures were not different than the artificial diet (control). Furthermore, chemical analysis of headspace samples of manures revealed qualitative differences in odor (volatile) profiles of all manures and artificial diet, indicating that behavioral differences could be due to the differences in the volatile chemistry of the adult ovipositional substrates and larval growth mediums. This study may contribute toward both understanding the linkage between ecological adaptations and host selection mechanisms and the development of pest management strategies against this serious pest of medical and veterinary importance.

The pig roundworm, Ascaris suum, is commonly found in domestic pigs all over the world. The transmission to humans takes place by ingestion of infective A. suum eggs present in soil because pig manure is widely used as fertilizer. The possible role of A. suum in the human visceral larva migrans (VLM) syndrome has been discussed controversially during past decades, even though various case reports, particularly from Japan document pulmonal, hepatic and even cerebral symptoms caused by migrating A. suum larvae after ingestion of infected row meat (liver) or contaminated vegetables. We examined 4481 sera by A. suum immunoblot (As-IB) and 5301 sera by Toxocara-ELISA from patients with symptoms associated with the VLM syndrome during three consecutive years (2012-2014). The incidence of A. suum-specific antibodies was 13.2 %, the incidence of T. canis specific antibodies 12.9 % and from a part of the As-IB positive sera (n = 417) additional Toxocara serology was performed to demonstrate the specificity of our tests. Only 56 out of the 417 (13.4 %) sera showed antibodies to both helminth species demonstrating that double infections exist. Interestingly the age distribution of the patients showed that 2.8 % of the Ascaris-positive patients were younger than 21 years, while in the Toxocara-positive group 13.4 % were <21 years. These results are in accordance with a Dutch study suspecting different ways of transmission as cause for this interesting age distribution. Due to the fact that large amounts of untreated pig manure are used as fertilizer and that the expulsion of adult A. suum worms causing intestinal ascariosis is extremely rare in Central European countries, the zoonotic potential of A. suum is considerably underestimated. We suggest that the performance of reliable immunoserological tests, in all industrialized countries where pigs are raised and their manure is used as fertilizer, could help to assess the actual potential of A. suum as causative agent of the VLM syndrome in humans.

Various separation technologies are used to deal with the enormous amounts of animal waste that large livestock operations generate. When the recycled waste stream is land applied, it is essential to lower the pathogen load to safeguard the health of livestock and humans. We investigated whether cationic polymers, used as a flocculent in the solid/liquid separation process, could reduce the pathogen indicator load in the animal waste stream. The effects of low charge density cationic polyacrylamide (CPAM) and high charge density cationic polydicyandiamide (PDCD) were investigated. Results demonstrated that CPAM was more effective than PDCD for manure coagulation and flocculation, while PDCD was more effective than CPAM in reducing the pathogen indicator loads. However, their combined use, CPAM followed by PDCD, resulted in both improved solids separation and pathogen indicator reduction., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: To evaluate the effect of schistosomiasis control in towns that had reached the criteria of infection controlled in Nanchang City., Methods: The residents and livestock of 14 towns that had reached the criteria of infection controlled in Nanchang, Xinjian and Jinxian counties of Nanchang City were sampled to investigate the schistosomiasis endemic situation, Oncomelania hupensis snail status, and data of schistosomiasis control work., Results: The positive rate of serum tests of the residents was 2.58%, and no positive stool samples were found. No cattle or sheep were positive in stool tests. Totally 1 635 snails were caught, and 1278 were alive with an average density of living snails of 0.051/0.1 M2, while no schistosome infected snails were found. All the surveyed administrative villages had the completed files of schistosomiasis control. Conclusion All the 14 towns that had reached the criteria of infection controlled of schistosomiasis have already reached the criteria of transmission controlled in Nanchang City.

Objective: To explore the existence of natural foci of Metorchis taiwanensis in Wuhu area., Methods: The ecological environment and daily living habits of residents were investigated; the eggs of M. taiwanensis in the dung of ducks in local region were collected by the sedimentation method; the Parafossarulus striatulus were disposed by the tabletting method and examined with a microscope to isolate the rediae and cercariae; the metacercariae in Pseudorasbora parve were isolated by using the homogenation. The duckling were fed directively by metacercaria of M. taiwanensis or by P. parve infected with metacercaria of M. taiwanensis for artificial infection, and M. taiwanensis were separated from ducks by dissecting the gallbladder and bile ducts., Results: The survey revealed that there were staggered lake river, rich vegetation, overgrown shrubs, aquatic plants, rich variety of freshwater snails and fish resources in Wuhu area, and it was home to a variety of waterfowl and duck. The average detection rate of eggs of M. tanwanensis in stool of ducks was 33.33% (10/30); that of rediae and cercarriae in P. striatulus was 1.17%(7/600); and that of encysted metacercaria in P. parve was 13.33% (8/60); the number of M. taiwanensis detected in the infected ducklings in 3 groups were 31, 8 and 0, respectively, the natural M. tanwanensis infection rate of ducks was 23.33% (7/30)., Conclusion: We have confirmed the existence of natural foci of M. tanwanensis in Wuhu area.

Manure from animal farms and sewage sludge contain pathogens and opportunistic organisms in various concentrations depending on the health of the herds and human sources. Other than for the presence of pathogens, these waste substances are excellent nutrient sources and constitute a preferred organic fertilizer. However, because of the pathogens, the risks of infection of animals or humans increase with the indiscriminate use of manure, especially liquid manure or sludge, for agriculture. This potential problem can increase with the global connectedness of animal herds fed imported feed grown on fields fertilized with local manures. This paper describes a simple, easy-to-use, low-tech hygienization method which conserves nutrients and does not require large investments in infrastructure. The proposed method uses the microbiotic shift during mesophilic fermentation of cow manure or sewage sludge during which gram-negative bacteria, enterococci and yeasts were inactivated below the detection limit of 3 log10 cfu/g while lactobacilli increased up to a thousand fold. Pathogens like Salmonella, Listeria monocytogenes, Staphylococcus aureus, E. coli EHEC O:157 and vegetative Clostridium perfringens were inactivated within 3 days of fermentation. In addition, ECBO-viruses and eggs of Ascaris suum were inactivated within 7 and 56 days, respectively. Compared to the mass lost through composting (15-57%), the loss of mass during fermentation (< 2.45%) is very low and provides strong economic and ecological benefits for this process. This method might be an acceptable hygienization method for developed as well as undeveloped countries, and could play a key role in public and animal health while safely closing the nutrient cycle by reducing the necessity of using energy-inefficient inorganic fertilizer for crop production.

There is limited knowledge about the survival of geohelminths, which are soil-transmitted human pathogens, in mesophilic anaerobic digestion processes. This study examined the fate of embryonated and unembryonated Ascaris suum ova in six laboratory-scale mesophilic (35 °C) anaerobic digesters processing swine manure to identify their survival strategies and investigate potential mechanisms to enhance their destruction. There was no significant difference in inactivation of Ascaris suum ova in digesters operated at different solids residence times (SRT) or feeding frequencies. Ova exposed to an anaerobic environment became dormant, or remained unembryonated throughout their residence in the reactors. Approximately 65% of ova were able to retain their viability for up to 16 days, after which the rate of inactivation increased until nearly all ova were nonviable by day 24. In contrast, ova exposed to aerobic conditions did not become dormant and progressed through several developmental stages until day 16, after which nearly all ova were observed to be nonviable. In addition, only 35% of fully developed ova exposed to the anaerobic environment retained their viability by day 16 compared to 65% for dormant ova. Results suggest that some ova are physically destroyed during digestion and ova can be inactivated faster if their development cycle is aerobically triggered before entering the anaerobic digestion process. Results also suggest that transfer of resource recovery technologies such as mesophilic anaerobic digestion to developing world settings must account for local climatic and health conditions so mutually beneficial outcomes can be attained.

House flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.), (Diptera: Muscidae), common pests on equine facilities, were studied in the laboratory to determine the success and duration of larval development and oviposition preferences on six substrates commonly found on equine facilities. Substrates tested were hay soiled with urine and manure, fresh horse manure, pine shaving bedding soiled with urine and manure (<12 h old), pine shaving bedding soiled with urine and manure (aged >72 h in a manure pile), builders sand bedding soiled with urine and manure aged 3 d, and soil from an overgrazed pasture mixed with urine and manure of variable age. House fly larvae failed to develop into adults in hay, soil, and sand substrates. Stable flies preferred to oviposit on substrates with plant material and not on fresh manure. However, when eggs were added to the substrates, pupariation was maximal in fresh manure and the fresh pine shaving substrate. Stable flies developed in all six equine substrates, but development was less successful on the substrates with soil. In choice tests, fresh manure and the fresh pine shaving substrates were the most attractive for house fly oviposition. These substrates also yielded the greatest number of house fly puparia from artificially added eggs. An understanding of oviposition preferences and differential larval development of house flies and stable flies on these substrates may help develop options for reducing pest populations by effectively managing equine waste and selecting appropriate bedding materials., (© 2014 Entomological Society of America.)

Animal waste from concentrated swine farms is widely considered to be a source of environmental pollution, and the introduction of veterinary antibiotics in animal manure to ecosystems is rapidly becoming a major public health concern. A housefly larvae (Musca domestica) vermireactor has been increasingly adopted for swine manure value-added bioconversion and pollution control, but few studies have investigated its efficiency on antibiotic attenuation during manure vermicomposting. In this study we explored the capacity and related attenuation mechanisms of antibiotic degradation and its linkage with waste reduction by field sampling during a typical cycle (6 days) of full-scale larvae manure vermicomposting. Nine antibiotics were dramatically removed during the 6-day vermicomposting process, including tetracyclines, sulfonamides, and fluoroquinolones. Of these, oxytetracycline and ciprofloxacin exhibited the greater reduction rate of 23.8 and 32.9 mg m(-2), respectively. Environmental temperature, pH, and total phosphorus were negatively linked to the level of residual antibiotics, while organic matter, total Kjeldahl nitrogen, microbial respiration intensity, and moisture exhibited a positive effect. Pyrosequencing data revealed that the dominant phyla related to Firmicutes, Bacteroidetes, and Proteobacteria accelerated manure biodegradation likely through enzyme catalytic reactions, which may enhance antibiotic attenuation during vermicomposting.

Eggs of Ascaris suum from pigs are highly resistant and commonly used as a conservative indicator of pathogen inactivation during slurry storage. Eggs of Ascaridia galli, the poultry ascarid, are also known to be highly resistant but the suitability as an indicator of pathogen inactivation has never been tested. Pig slurry has to be stored for several months to inactivate pathogens but chemical treatment of slurry may reduce this time. The suitability of A. galli as an indicator of slurry sanitation was tested by comparing the survival of eggs of A. suum and A. galli in pig slurry. In addition, the effect of urea treatment on inactivation of ascarid eggs in relation to storage time was also tested. Nylon bags with 10,000 eggs of either species were placed in 200 ml plastic bottles containing either urea-treated (2%) or untreated pig slurry for up to 120 days at 20°C, 6 days at 30°C, 36h at 40°C or 2h at 50°C. At all the temperatures in both slurry types, A. galli eggs were inactivated at a significantly faster rate (P<0.05) compared to A. suum eggs. For each 10°C raise in temperature from 20°C, T50 (time needed to inactivate 50% of eggs) for both types of eggs was reduced markedly. At all temperatures, viability of eggs of both species was significantly higher (P<0.05) in untreated slurry compared to urea-treated slurry except A. galli eggs at 20°C where no significant difference was detected. In untreated slurry, the levels of pH (6.33-9.08) and ammonia (0.01-1.74 mM) were lower (P<0.0001) compared to that of urea-treated slurry (pH: 8.33-9.28 and ammonia 1-13 mM). The study demonstrated that A. galli eggs are more sensitive to unfavourable conditions compared to A. suum eggs. The use of A. galli eggs as hygiene indicator may thus be suitable to assess inactivation of pathogens that are more sensitive than A. galli eggs. Addition of urea may markedly reduce the storage time of slurry needed to inactivate A. suum and A. galli eggs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Earthworms can accumulate heavy metals in their intestines to a great extent. Impact of feed materials and duration of metal exposure on natural activity of earthworms are rather unclear; this investigation therefore addresses the impact of metal rich Tea Factory Coal Ash (TFCA) on reproduction, composting and metal accumulation ability of Eisenia fetida and Lampito mauritii. Earthworm count and cocoon production increased significantly during vermicomposting. pH of the vermicomposted mixtures shifted toward neutrality, total organic C decreased substantially and total N enhanced significantly compared to composting. High heavy metal (Mn, Zn, Cu, As) accumulation was recorded in the intestine of both the earthworm species. Moreover, gradual increase in the metal-inducible metallothionein concentration indicated the causal mechanism of metal accumulation in these species. TFCA+cow dung (CD) (1:1) were most favorable feed mixture for E. fetida and TFCA+CD (1:2) were good for L. mauritii in regard to metal accumulation and compost quality., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Two rates (0.4 mg/kg body weight/d and 0.6 mg/kg body weight/d) of a daily feed-through formulation of novaluron (Novaluron 0.67% active ingredient Cattle Mix), a newer benzoylphenyl urea insecticide, were evaluated for efficacy in controlling the larval stage of horn flies, Haematobia irritans (L.), house flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.), developing in cow manure. Both rates of feed-through novaluron, delivered consecutively for 10 d, reduced adult emergence of all three species when compared with the untreated control. The presence of deformed puparia indicated that novaluron had an insect growth regulator effect on the developing fly larvae. Both of the feed-through rates evaluated resulted in 100% reduction of adult stable fly emergence after the second day of feed-through treatment. The level of control efficacy observed against these three fly species make this feed-through formulation a candidate for use in an integrated livestock pest management program, particularly in confined cattle production situations where a feed-through product could be easily administered.

Only a few aspects of the biology of Amblyomma parvitarsum Neumann are known. Adults of this hard tick species are parasites of South American camelids in the Andean plateau of Argentina, Bolivia, Peru and Chile and also in the Argentine Patagonia, but they have been also rarely found on other artyodactils and two species of birds. The larva has been collected from reptiles in northern Chile, whereas the hosts for the nymph remain unknown. On nine localities included within Altitude Tropical and Perarid Mediterranean ecoregions in northern Chile, we analyzed 237 reptiles, 285 birds, 624 rodents and 52 camelids for infestation with A. parvitarsum to calculate seasonal prevalence of this tick. We also reviewed the literature of this tick and three entomological collections for obtaining and summarizing all the information to date about this tick. None of the analyzed birds and rodents were parasitized with A. parvitarsum; however, seven over a total of ten reptile species that we caught were infested with the larva. In the camelid species Lama glama and Vicugna pacos we collected adult specimens of this tick. Larval prevalence was higher during fall (75 %) in Liolaemus pleopholis in the Altitude Tropical ecoregion. We also collected adult specimens of A. parvitarsum from camelid manure heaps during summer in Salar de Surire and Llullaillaco localities. Additionally, we also reviewed the literature of this tick and examined specimens in three entomological collections for obtaining and summarizing all the information to date about this tick. By this study, nine localities and seven new hosts are added for A. parvitarsum and we confirm reptiles as specific hosts of this tick larva.

Insects feeding on plant sap, blood, and other nutritionally incomplete diets are typically associated with mutualistic bacteria that supplement missing nutrients. Herbivorous mammal dung contains more than 86% cellulose and lacks amino acids essential for insect development and reproduction. Yet one of the most ecologically necessary and evolutionarily successful groups of beetles, the dung beetles (Scarabaeinae) feeds primarily, or exclusively, on dung. These associations suggest that dung beetles may benefit from mutualistic bacteria that provide nutrients missing from dung. The nesting behaviors of the female parent and the feeding behaviors of the larvae suggest that a microbiome could be vertically transmitted from the parental female to her offspring through the brood ball. Using sterile rearing and a combination of molecular and culture-based techniques, we examine transmission of the microbiome in the bull-headed dung beetle, Onthophagus taurus. Beetles were reared on autoclaved dung and the microbiome was characterized across development. A ~1425 bp region of the 16S rRNA identified Pseudomonadaceae, Enterobacteriaceae, and Comamonadaceae as the most common bacterial families across all life stages and populations, including cultured isolates from the 3(rd) instar digestive system. Finer level phylotyping analyses based on lepA and gyrB amplicons of cultured isolates placed the isolates closest to Enterobacter cloacae, Providencia stuartii, Pusillimonas sp., Pedobacter heparinus, and Lysinibacillus sphaericus. Scanning electron micrographs of brood balls constructed from sterile dung reveals secretions and microbes only in the chamber the female prepares for the egg. The use of autoclaved dung for rearing, the presence of microbes in the brood ball and offspring, and identical 16S rRNA sequences in both parent and offspring suggests that the O. taurus female parent transmits specific microbiome members to her offspring through the brood chamber. The transmission of the dung beetle microbiome highlights the maintenance and likely importance of this newly-characterized bacterial community.

Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

Determining the presence of viable Cryptosporidium parvum oocysts in complex environmental matrices in hygiene control can prevent the contamination of water resources and food with this pathogen. This study assessed the induction ratio of hsp70 mRNA production by heat shock in different oocysts as a marker of viability. Using different procedures for (m)RNA extraction directly from manure and reverse transcription real-time qPCR, this study found slightly increased hsp70 mRNA contents in viable oocysts that were heat shock induced at 45°C for 20 min compared to not induced oocysts (1.6 fold induction in average). Prolonging the heat shock treatment to 2h did not further increase the copy numbers. Heat shock by consecutive stimuli, such as freezing and then heating, did not yield significantly higher copy numbers than the 45°C treatment. There was a certain background level of hsp70 mRNA in viable oocysts that were not exposed to heat shock, indicating a constitutive production of the transcripts in the oocysts. The production of hsp70 mRNA induced by heat shock in oocysts aged for 9 months that exhibited reduced viability was lower than in fresher oocysts (induction ratio<1.2). No production of hsp70 mRNA by heat shock was detected in 12 months old oocysts that were not viable in the excystation test. Oocysts inactivated at 75°C for 30 min were not able to respond to heat shock, and low amount of copies were occasionally measured only in total RNA extracts, but not in mRNA extracts that were purified directly with an oligo (dT)25 based system. The induction ratio of hsp70 mRNA varied according to the viability of the organisms in a sample. Copy numbers of β-tubulin mRNA in viable oocysts were lower than hsp70 mRNA, therefore the latter is more suitable to detect low numbers of oocysts by RT-qPCR., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The protection of drinking water from pathogens such as Cryptosporidium and Giardia requires an understanding of the short-term microbial release from faecal contamination sources in the catchment. Flow-weighted samples were collected during two rainfall events in a stream draining an area with on-site sewers and during two rainfall events in surface runoff from a bovine cattle pasture. Samples were analysed for human (BacH) and ruminant (BacR) Bacteroidales genetic markers through quantitative polymerase chain reaction (qPCR) and for sorbitol-fermenting bifidobacteria through culturing as a complement to traditional faecal indicator bacteria, somatic coliphages and the parasitic protozoa Cryptosporidium spp. and Giardia spp. analysed by standard methods. Significant positive correlations were observed between BacH, Escherichia coli, intestinal enterococci, sulphite-reducing Clostridia, turbidity, conductivity and UV254 in the stream contaminated by on-site sewers. For the cattle pasture, no correlation was found between any of the genetic markers and the other parameters. Although parasitic protozoa were not detected, the analysis for genetic markers provided baseline data on the short-term faecal contamination due to these potential sources of parasites. Background levels of BacH and BacR makers in soil emphasise the need to including soil reference samples in qPCR-based analyses for Bacteroidales genetic markers.

Application of manure or soil amendments of animal origin (untreated soil amendments; UTSAs) to agricultural land has been a long-standing practice to maintain or improve soil quality through addition of organic matter, nitrogen, and phosphorus. Much smaller quantities of these types of UTSAs are applied to land used for food crops than to land used for animal grain and forage. UTSAs can harbor zoonotic enteric pathogens that may survive for extended periods after application. Additional studies are needed to enhance our understanding of preharvest microbial food safety hazards and control measures pertaining to the application of UTSAs especially for land used to grow produce that may be consumed raw. This document is intended to provide an approach to study design and a framework for defining the scope and type of data required. This document also provides a tool for evaluating the strength of existing data and thus can aid the produce industry and regulatory authorities in identifying additional research needs. Ultimately, this framework provides a means by which researchers can increase consistency among and between studies and facilitates direct comparison of hazards and efficacy of controls applied to different regions, conditions, and practices.

Because of outbreaks of cryptosporidiosis in humans, some Cryptosporidium spp. have become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by Cryptosporidium suis , a fraction may be comprised of other Cryptosporidium species infectious to humans such as Cryptosporidium parvum . To better understand the survival dynamics of Cryptosporidium spp., oocysts associated with swine operations, 2 experiments were performed to determine die-off rates of C. parvum oocysts in a swine waste lagoon (2009 and 2010) and its spray field (2010 and 2011). Sentinel chambers containing a lagoon effluent suspension of C. parvum oocysts were submerged in the lagoon, and triplicate chambers were removed over time; oocysts were extracted and assayed for viability. For comparative purposes, inactivation rates of Ascaris suum eggs contained in sentinel chambers were also determined. For 2 spray field experiments, air-dried and sieved surface soil was placed in sentinel chambers, hydrated, and inoculated with a lagoon effluent suspension of C. parvum oocysts. Sentinel chambers and control oocysts in PBS contained in microcentrifuge tubes were buried 1.5 cm below the soil surface in 3 blocks. Triplicate chambers and controls were removed over time; oocysts were extracted and assayed for viability. Based on the first order decay equation, days to reach 99% die-off (T(99)) were determined. T(99)-values determined for the 2 lagoon experiments were 13.1 and 20.1 wk, respectively. A T(99)-value for C. parvum in the spray field was significantly longer at 38.0 wk than the control oocysts in PBS at 29.0 wk. The waste lagoon and spray field system of manure management at this large-scale farrowing operation appeared to reduce the load of C. parvum oocysts before they can be hydrologically transported off the operation and reduces their likelihood of contaminating surface waters and threatening public health.

Separation of pig slurry into solid and liquid fractions is gaining importance as a way to manage increasing volumes of slurry. In contrast to solid manure and slurry, little is known about pathogen survival in separated liquid slurry. The viability of Ascaris suum eggs, a conservative indicator of fecal pollution, and its association with ammonia was investigated in separated liquid slurry in comparison with raw slurry. For this purpose nylon bags with 6000 eggs each were placed in 1 litre bottles containing one of the two fractions for 308 days at 5 °C or 25 °C. Initial analysis of helminth eggs in the separated liquid slurry revealed 47 Ascaris eggs per gramme. At 25 °C, egg viability declined to zero with a similar trend in both raw slurry and the separated liquid slurry by day 308, a time when at 5 °C 88% and 42% of the eggs were still viable in separated liquid slurry and raw slurry, respectively. The poorer survival at 25 °C was correlated with high ammonia contents in the range of 7.9-22.4 mM in raw slurry and 7.3-23.2 mM in liquid slurry compared to 3.2-9.5 mM in raw slurry and 2.6-9.5 mM in liquid slurry stored at 5 °C. The study demonstrates that at 5 °C, A. suum eggs have a higher viability in separated liquid slurry as compared to raw slurry. The hygiene aspect of this needs to be further investigated when separated liquid slurry is used to fertilize pastures or crops.

The Cu, Ni and Zn accumulations in leaves and roots of lettuce (Lactuca sativa L) grown in soil amended with natural and contaminated cattle manure vermicompost were evaluated. The vermicompost residues containing relatively high metal concentrations used in this work were obtained from a previous experiment, in which vermicompost was applied to removing metals from electroplating wastes. Sequential lettuce cultivations were conducted in pots containing the residual substrates from the first cultivation by adding metal-enriched vermicompost residues. In general, the Cu, Ni and Zn concentrations in leaves and roots of lettuce plants grown in vermicompost enriched with these metals were higher than in the treatment using the natural vermicompost. The metal concentrations in leaves from treatments with natural vermicompost were below the critical concentrations of toxicity to plants. However, the metal concentrations in leaves of the third cultivation in which metal-enriched vermicompost was applied were greater than the upper limit that causes plant toxicity, but no visual damage was observed in the plants. Treatment with Zn-enriched vermicompost resulted in toxicity symptoms, but plant damage did not result in the death of the plant. The chemical fractionation of Cu, Ni and Zn in residues from lettuce cultivation was evaluated by using a sequential extraction procedure and metal concentrations were increased in the different chemical fractions according to the increase of vermicompost dose.

Culicoides species (Diptera: Ceratopogonidae) belonging to the Obsoletus and Pulicaris groups are considered to be the main vectors of bluetongue virus (BTV) in non Mediterranean Europe. Selected terrestrial microhabitats (n=17) on a farm in northern Spain were sampled repeatedly over a year-long period and characterized for use by Culicoides species for immature development. Concurrent use of CDC light traps showed the presence of 37 species and 66,575 specimens of adult Culicoides. A total of 28 species and 11,396 individuals emerged from laboratory-maintained soil samples. Culicoides obsoletus and Culicoides scoticus (pooled as Obsoletus complex) were particularly abundant (comprising 58.6% and 74.5% of the total collections in light traps and emergence traps respectively). Potential key vectors of animal viruses (such as BTV) were found in two main terrestrial types of microhabitats. In the case of C. obsoletus, different types of manure (old and composted manure, manure mixed with organic matter, and fresh manure) produced most of the specimens. In contrast, larvae of C. scoticus and Culicoides lupicaris were associated with soil substantially comprised of rotting leaf litter that included the parasitic plant Lathraea clandestina. Several species, Culicoides festivipennis, Culicoides punctatus and Culicoides brunnicans, were very common in mud at pond margins. Indeed, pond microhabitats and runoff below barn rooflines supported the greatest species richness. In the pond habitat, 49.4% of Culicoides specimens emerged from mud at the water edge, as opposed to 50 cm above (32.4%) and 1 meter above waterline (18%). Similar species richness, but statistically significant differences in abundance, were observed among the four pond microhabitats. Overall, the majority of the specimens were found in the upper layer (0-3 cm), except in manure, where they preferred deeper layers (>6 cm). Maximum peaks of abundance occurred in both light traps and soil samples in summer months, whereas increased captures in autumn were noticed only in light traps. Both trapping systems failed to collect adult Culicoides midges in the coldest months of December, January and February. The literature on immature habitats of species suspected in BTV transmission in Europe, the Pulicaris group and particularly the Obsoletus group, is briefly reviewed., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The protozoan parasite Cryptosporidium is a major cause of diarrhoea worldwide. This paper presents the first model-based inventory with 0.5 by 0.5 degree resolution of global Cryptosporidium emissions for the year 2000 from humans and animals to surface water. The model is based on nutrient distribution modelling, because the sources and transport of oocysts and nutrients to the surface water are comparable. Total emissions consist of point source emissions from wastewater and nonpoint source emissions by runoff of oocysts in manure from agricultural lands. Results indicate a global emission of 3 × 10(17) oocysts per year, with comparable contributions from point and nonpoint sources. Hot-spot areas for point sources are big cities in China, India and Latin America, while the area with the largest nonpoint source emissions is in China. Uncertainties in the model are large. Main areas for further study are (i) excretion rates of oocysts by humans and animals, (ii) emissions of humans not connected to sewage systems, and (iii) retention of oocysts to determine surface water pathogen concentrations rather than emissions. Our results are useful to health organisations to identify priority areas for further study and intervention., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In the present study the potential of the earthworm Eisenia fetida to process large amounts of waste was evaluated through continuous feeding reactors in which new layers of rabbit manure were added sequentially to form an age gradient inside the reactors. An optimal moisture level, ranging from 66% to 76%, was maintained throughout the process using an automatic watering system. The pH was close to 8.3, but decreased to 7.6 after 200 d of vermicomposting. No changes in electrical conductivity through the profile of layers were detected. Based on comparisons of phospholipid fatty acid (PLFA) profiles and microbial activity measurements (basal respiration), a decrease in the levels of bacteria and fungi in layers corresponding to vermicomposting times of more than 200 d occurred. This points to a higher degree of stabilisation in the final product, which is of utmost importance for its safe use as an organic amendment., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The house fly, Musca domestica L. (Diptera: Muscidae) is one of the major pests of confined and pastured livestock worldwide. Livestock manures play an important role in the development and spread of M. domestica. In the present study, we investigated the impact of different livestock manures on the fitness and relative growth rate of M. domestica and intrinsic rate of natural increase. We tested the hypotheses by studying life history parameters including developmental time from egg to adult's eclosion, fecundity, longevity, and survival on manures of buffalo, cow, nursing calf, dog, horse, poultry, sheep, and goat, which revealed significant differences that might be associated with fitness costs. The maggots reared on poultry manure developed faster compared to any other host manure. The total developmental time was the shortest on poultry manure and the longest on horse manure. The fecundity by females reared on poultry, nursing calf, and dog manures was greater than on any other host manures. Similarly, percent survival of immature stages, pupal weight, eggs viability, adults' eclosion, survival and longevity, intrinsic rate of natural increase, and biotic potential were significantly higher on poultry, nursing calf, and dog manures compared to any other livestock manures tested. However, the sex ratio of adult flies remained the same on all types of manures. The low survival on horse, buffalo, cow, sheep, and goat manures suggest unsuitability of these manures, while the higher pupal weight on poultry, nursing calf, and dog manures suggest that these may provide better food quality to M. domestica compared with any other host manures. Our results point to the role of livestock manures in increasing local M. domestica populations. Such results could help to design cultural management strategies which may include sanitation, moisture management, and manure removal.

Spinosad, diatomaceous earth, and cyfluthrin were assessed on two broiler farms at Gleneagle and Gatton in southeastern Queensland, Australia in 2004-2005 and 2007-2009, respectively to determine their effectiveness in controlling lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Insecticide treatments were applied mostly to earth or 'hard' cement floors of broiler houses before the placement of new bedding. Efficacy of each agent was assessed by regular sampling of litter and counting of immature stages and adult beetles, and comparing insect counts in treatments to counts in untreated houses. Generally, the lowest numbers of lesser mealworm were recorded in the house with hard floors, these numbers equalling the most effective spinosad applications. The most effective treatment was a strategic application of spinosad under feed supply lines on a hard floor. In compacted earth floor houses, mean numbers of lesser mealworms for two under-feed-line spinosad treatments (i.e., 2-m-wide application at 0.18 g of active insecticide (g [AI]) in 100-ml water/m2, and 1-m-wide application at 0.11 g ([AI] in 33-ml water/m2), and an entire floor spinosad treatment (0.07 g [AI] in 86-ml water/m2) were significantly lower (i.e., better control) than those numbers for cyfluthrin, and no treatment (controls). The 1-m-wide under-feed-line treatment was the most cost-effective dose, providing similar control to the other two most effective spinosad treatments, but using less than half the active component per broiler house. No efficacy was demonstrated when spinosad was applied to the surface of bedding in relatively large volumes of water. All applications of diatomaceous earth, applied with and without spinosad, and cyfluthrin at the label rate of 0.02 g (AI)/100-ml water/m2 showed no effect, with insect counts not significantly different to untreated controls. Overall, the results of this field assessment indicate that cyfluthrin (the Australian industry standard) and diatomaceous earth were ineffective on these two farms and that spinosad can be a viable alternative for broiler house use.

A granular formulation of novaluron (Novaluron 0.2G, 0.2% [AI]), a newer benzoylphenyl urea insecticide, was evaluated for its efficacy in controlling the larval stage of horn flies, Haematobia irritans (L.); house flies, Musca domestica L.; and stable flies, Stomoxys calcitrans (L.), in cow manure. Various rates and insecticide placement locations (top, middle, and bottom of manure) were evaluated in this study and all combinations of these variables reduced adult emergence of all three species when compared with the untreated controls. The presence of deformed pupae indicated that novaluron had an insect growth regulator effect on the developing fly larvae. Top, middle, or bottom application rates of 0.125, 0.195, 0.25, and 0.375 g novaluron onto manure samples, reduced adult horn fly emergence by > 90%. Middle and bottom application rates of 0.195, 0.25, and 0.375 g novaluron reduced adult house fly emergence >93%. All rates and placement combinations resulted in >98% reduction of adult stable fly emergence. The level of control efficacy observed against these three fly species along with the ease of use of a granular formulation, make this product an ideal candidate for use in an integrated livestock pest management program.

Giardia duodenalis is a protozoan parasite known to infect animals and humans. Zoonotic transmission of G. duodenalis can occur by the consumption of drinking water produced from surface water that is contaminated by runoff from manure-laden fields or pastures. Although it was previously reported that storing solid cattle manure decreases G. duodenalis cyst viability, no data are available on cyst survival in slurry waste from cattle. In this study the number and the viability of G. duodenalis cysts was determined in cattle slurry for up to 90 days. G. duodenalis cysts were counted in 30 slurry samples with a quantitative direct immunofluorescence assay. The geometric mean number of cysts was reduced by 77% after 90 days (P<0.0014), although there was substantial variability between samples. A fluorogenic dye staining using 4',6'-di-amino-2-phenylindole and propidium iodide showed a decreased viability from 45 days onwards, and after 90 days incubation, only 3% of the cysts were viable. Gerbils and lambs were artificially infected with 50 day-old and 90 day-old cysts and faecal excretion of G. duodenalis was monitored between 5 and 7 days after infection. Seven days after infection the gerbils were euthanized for Giardia trophozoite counts. Although one cyst was found in the faeces of one of the gerbils after infection with 50 day-old cysts, no trophozoites were recovered from the intestines of any gerbil (n=8). Experimental infection of lambs with 10(5)50 day-old and 90 day-old slurry cysts caused low cyst excretion in one out of two and one out of three lambs, respectively. Together, these data show that storage of cattle slurry for 90 days greatly reduces the number and viability of G. duodenalis cysts., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Cryptosporidium and Giardia (oo)cyst concentrations are frequently used for assessing drinking water safety. The widely used USEPA Method 1623 provides total counts of (oo)cysts, but may not be accurate for human health risk characterization, since it does not provide infectivity information. The total counts and infectious fraction of Cryptosporidium oocysts and the total counts of Giardia cysts were assessed in major fecal pollution sources. Fresh calf and cow feces, their manure, and the discharge point were sampled in a small rural sub-watershed (n = 20, 21, 10, 10). Median concentrations for total (oo)cysts were higher in calves (333 oocysts g(-1); 111 cysts g(-1)) than in cows (52 oocysts g(-1); 7 cysts g(-1)). Infectious oocysts were found in 17 (7%) of the samples and none were found in manure or at the discharge point. Urban sources were sampled in the influent and effluent (n = 19, 18) of two wastewater treatment plants. Peak concentrations were 533 oocysts L(-1) and 9,010 cysts L(-1) for influents and 89 oocysts L(-1) and 472 cysts L(-1) for effluents. Infectious oocyst fractions varied from below the detection limit to 7-22% in the effluent and influent respectively. These infectious fractions are significantly lower than those currently used for quantitative microbial risk assessment estimates.

As part of the C-EnterNet surveillance program of the Public Health Agency of Canada, 122 pooled swine manure samples from 10 farms in Ontario, Canada were collected and tested for Giardia and Cryptosporidium. Giardia duodenalis cysts and Cryptosporidium spp. oocysts were detected using immunofluorescence microscopy. Nested-polymerase chain reaction protocols were performed to amplify the small subunit rRNA gene and the β-giardin gene for G. duodenalis, and the small subunit rRNA gene and the heat shock protein-70 gene for Cryptosporidium spp. The DNA amplicons were sequenced to determine genotypes and species. A mixed multivariable method was used to compare the presence of Giardia and Cryptosporidium in different stages of production. Both Giardia cysts and Cryptosporidium oocysts were present on all tested farms, with 50.8% of the samples positive for G. duodenalis and 44.3% positive for Cryptosporidium spp. by microscopy, and 66.4% and 55.7%, respectively, positive by polymerase chain reaction (PCR). No significant agreement was observed between microscopy and PCR method to detect Giardia and Cryptosporidium (p<0.05). The prevalence of Giardia in manure pits and finisher pigs did not differ (p>0.05), however, it was less frequent (odds ratio, OR=0.21 [0.07, 0.63]) among sows. Cryptosporidium was more likely (OR=3.6 [1.3, 9.9]) to be detected in manure pits and weaners (OR=3.3 [1.1, 10.0]) compared to finisher pigs, and it was less frequent (OR=0.06 [0.007, 0.55]) in sows than in finishers (p<0.05). DNA sequencing demonstrated that 92.1% of the Giardia isolates were Assemblage B and 7.9% were Assemblage E. The most prevalent Cryptosporidium were Cryptosporidium parvum (55.4%), and Cryptosporidium sp. pig genotype II (37.5%). These findings indicate that the occurrence of zoonotic isolates of G. duodenalis and Cryptosporidium is very high on swine farms in southern Ontario, and that there is a potential for transmission between swine and humans by means of cyst and oocyst contaminated water or foods.

This paper reports the recycling of nutrients by vermicomposting of cow dung (CD), poultry droppings (PD) and food industry sludge (FIS) employing earthworms (Eisenia fetida). A total of six vermicomposting units were established and dynamics of chemical and biological parameters has been studied for 13 weeks. The waste mixture containing 50% CD+25% PD+25% FIS had better fertilizer value among studied waste combinations. At the end of experiment, vermicomposts showed decrease in pH and organic C, but increase in EC, total Kjeldhal N, total available P and total K contents. The C:N ratio of final vermicomposts also reduced to 10.7-12.7 from 22.8 to 56 in different waste combinations. The earthworms have good biomass gain and cocoon production in all vermicomposting units but CD alone and 50% CD+25% PD+25% FIS were better than other studied combinations., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The objective of this study was to compare the occurrence and the genotypes and species of Giardia duodenalis and Cryptosporidium spp. in beef and dairy cattle from farms in the Regional Municipality of Waterloo, Ontario, in an effort to determine the potential for zoonotic transmission from these animals. Pooled manure samples were collected from 45 dairy cattle farms and 30 beef cattle farms. The presence of Giardia cysts and Cryptosporidium oocysts was determined by immunofluorescence microscopy, while nested-PCR and DNA sequencing were used to determine genotypes and species. The overall farm prevalence was very high for both Giardia and Cryptosporidium, and was similar for dairy cattle farms (96 and 64%, respectively) and beef cattle farms (97 and 63%, respectively). However, on dairy cattle farms, G. duodenalis and Cryptosporidium spp. were detected in 44% and 6% of total pooled pen manure samples, respectively, with the occurrence of both parasites being generally higher in calves than in older animals. Most Giardia isolates were identified as either the host-adapted genotype G. duodenalis Assemblage E or the zoonotic Assemblage B. Cryptosporidium parvum and Cryptosporidium andersoni were the most frequently identified species in dairy cattle, while the non-zoonotic species Cryptosporidium ryanae and Cryptosporidium bovis were also found. On beef cattle farms, 72% and 27% of the total pooled pen manure samples were positive for Giardia and Cryptosporidium, respectively, with no obvious correlation with age. All Giardia isolates in beef cattle were identified as G. duodenalis Assemblage E, while all Cryptosporidium isolates were identified by sequence analysis as C. andersoni, although microscopic analyses, and subsequent restriction fragment length polymorphism analyses, indicated that other Cryptosporidium species were also present. The results of this study indicate that although Giardia and Cryptosporidium were identified in a higher overall percentage of the pooled beef cattle manure samples than in dairy cattle, firmly established zoonotic genotypes and species were much more common in dairy cattle than in beef cattle in this region. Dairy cattle, and especially dairy calves, may, therefore, pose a greater risk of infection to humans than beef cattle. However, these results may also provide evidence of potential zooanthroponotic transmission (human to animal)., (Crown Copyright © 2010. Published by Elsevier B.V. All rights reserved.)

Metam sodium (MS, sodium N-methyldithiocarbamate) is a widely used soil pesticide. Fumigation or chemical sterilization of poultry litter containing infectious oocysts could be an effective strategy to block the transmission of avian coccidia. In the current study, the effect of MS on the viability and infectivity of ocysts was investigated. The development of isolated, unsporulated oocysts of both Eimeria tenella and Eimeria maxima was inhibited, in a dose-related manner (IC(50) 8 to 14 microg/ml), by exposure to aqueous MS. Most treated oocysts failed to develop beyond early stages of sporulation. To determine the effect of MS on infectivity, isolated oocysts of E. tenella , Eimeria acervulina , and E. maxima were exposed for 24 hr to aqueous concentrations of MS ranging from 0 to 1,000 microg/ml. Treated oocysts were inoculated into chickens, and parameters of coccidiosis infection were compared to chickens inoculated with equal numbers of untreated oocysts. In a dose-related manner, MS significantly reduced the infectivity of oocysts with maximum effect observed at a dose of 300 microg/ml. When a mixture of oocysts containing 3 coccidian species was exposed to 300 microg/ml MS, from 0 to 24 hr, infectivity of oocysts was significantly reduced after a minimum of 12 hr of exposure. Treatment of aqueous slurries of litter samples obtained from commercial poultry houses, with 300 microg/ml MS for 24 hr, prevented the sporulation of eimerian oocysts in the litter samples relative to untreated control samples. The results indicate that MS could be used to reduce coccidial contamination of poultry litter.

Three experiments were conducted in a University of Jordan field in the Al-Jubeiha area, to evaluate the effect of manure composting for 0, 2, 4, or 6 weeks prior to planting on the house fly (Musca domestica L.) population. The obtained results indicated that the housefly prefers broiler and layer manures over other tested materials for breeding. Composting can reduce house fly populations, especially with longer composting periods. Reduction in house fly numbers was linearly correlated with composting period. Even though organic manures composted for longer periods reduce house fly populations, broiler and layer manures were still favored.

Members of the public are always somewhat aware of foodborne and other zoonotic pathogens; however, recent illnesses traced to produce and the emergence of pandemic H1N1 influenza virus have increased the scrutiny on all areas of food production. The Council for Agricultural Science and Technology has recently published a comprehensive review of the fate and transport of zoonotic pathogens that can be associated with swine manure. The majority of microbes in swine manure are not zoonotic, but several bacterial, viral, and parasitic pathogens have been detected. Awareness of the potential zoonotic pathogens in swine manure and how treatment, storage, and handling affect their survival and their potential to persist in the environment is critical to ensure that producers and consumers are not at risk. This review discusses the primary zoonotic pathogens associated with swine manure, including bacteria, viruses, and parasites, as well as their fate and transport. Because the ecology of microbes in swine waste is still poorly described, several recommendations for future research are made to better understand and reduce human health risks. These recommendations include examination of environmental and ecological conditions that contribute to off-farm transport and development of quantitative risk assessments.