Profiling partner presence and abundance alongside plant host nutrients may highlight specific roles, if any, for microbes in an insect that lost endosymbionts. The patterns in AAT transcripts within the Phylloxeridae, and among galling and free-living Aphidomorpha, provide insight into the selection imparted through host manipulation and the evolution of endosymbiosis. Slimfast paralogs appear across many phylloxerids, and thus appear not to drive endosymbiosis. Rather the presence of slimfast duplications in free-living Phylloxera suggest an additional role in nutrient transport when the host cannot be manipulated. Although our data support the slimfast expansion among aphidomorphs, we hypothesize that plant host nutrient availability may have facilitated some duplications found in free-living pea and green peach aphids because several paralogs appear absent from galling aphids. Recent cellular localization screens indicated one of these genes increases in expression prior to and after Buchnera transmission as the bacteriocyte develops. Because galling aphids lack ACYPI008904 paralogs, selection to duplicate specific nutrient transporters may be relaxed when plant host nutrient status can be manipulated. Notably, square pots for planting the emergence of phylloxerid-specific duplications related to these aphid genes, highlights a bacteriocyte-independent role for some slimfast orthologs.
Plant pathogens have significant impacts on food production. In a time of intensive exchange of different planting material on national and international levels, data about distribution and economic importance of plant pathogens and plant diseases are crucial for their successful management. To decrease the negative impact that pathogens have, it is crucial to constantly work on their detection and geospatial distribution. Grapevine is an important plant crop with worldwide distribution. Throughout history, grapevines have been faced with challenges that changed European viticultural practice. Phylloxera , a pest that was introduced in Europe from America in the 19th century, caused the “Great French Wine Blight” . The use of American rootstocks solved the phylloxera problem, but resulted in the introduction of two fungal diseases: powdery and downy mildew . Demand for American rootstocks and priority given to grapevine varieties more resistant to fungal diseases resulted in dissemination of their planting material between continents and countries. Besides that, in many grape growing regions priority given to worldwide-grown cultivars , promoted by wine companies and markets, caused irretrievable loss of autochthonous cultivars . Intensive exchange of planting material opened the space for dissemination of grapevine viruses, at the time a group of practically unknown pathogens . Grapevines have 65 reported viruses, the highest number known from a single crop . However, only a fraction of these viral species is considered economically important. Because virus infection impacts crop quality and yield, data about virus distribution and frequency are important in the implementation of appropriate disease management practices.
So far, measures for virus control in vineyards are mainly based on control of vectors , use of virus-free planting material, and constant work on clonal and sanitary selection . For example, a California North Coast study on Grapevine leafroll-associated virus 3 showed benefits of using certified plant material of $0.40 per vine, $533 per acre, and $52.7 million per year for the region . In Croatia viticulture has a long tradition dating back to the Bronze Age . Today, approximately 15% of the ˇ Croatian population is associated with viticulture . Croatia has two different viticultural regions: continental, with continental temperate climate; and coastal, with influence of the Adriatic Sea and Mediterranean climate. In addition to popular cultivars , there are a significant number of autochthonous cultivars, primarily grown in coastal region. According to Maletic et al. there are at least ´ 125 autochthonous grapevine genotypes that represent valuable national heritage. The first report of grapevine viruses in Croatia included information on the spread and detrimental effect of viruses from infective degeneration complex . Viruses from the grapevine leaf- ´ roll complex were reported three decades later . Prevalence of GLRaV-3 in the coastal region has been documented , revealing infection of commercial vineyards and mixed infections with Grapevine virus A and Grapevine fleck virus. In the continental region, up to 52% vines surveyed were free of economically important viruses, with Grapevine leafroll-associated virus 1 and GFkV as the most common viruses . To improve the quality of ˇ planting material and save endangered autochthonous cultivars from extinction, clonal and sanitary selection programs were initiated 10 years ago .
The aim of this survey was to obtain additional knowledge about viruses present in autochthonous Croatian grapevine cultivars grown along the coastal region. Because of already confirmed prevalence of GLRaV-3, special attention was given to spatial distribution of its different variant groups. Results provide information that can be used for improving the quality of planting material and implementation of appropriate control measures.An adverse food reaction is defined as any abnormal clinical response that occurs following ingestion of a food or food component . It is often unclear if the pathologic mechanisms of AFR represent a specific immune-mediated response to food antigens, or if the mechanism is related to non-immune intolerance to a component in the food . When manifested as dermatological signs, an AFR is termed a cutaneous adverse food reaction . In addition to cutaneous signs, gastrointestinal signs, symmetric lupoid onychodystrophy, conjunctivitis, sneezing, and anaphylaxis have been associated with AFR in dogs; gastrointestinal and respiratory signs, conjunctivitis, and hyperactive behavior have also been reported in cats . Of these clinical signs, diarrhea and frequent defecation were most often diet-responsive in dogs; in the cat, diet-responsive clinical signs included vomiting and diarrhea . Among dogs and cats presented to their veterinarian for pruritus, the median prevalence of CAFR is estimated to be between 15% and 20% . In dogs in Australia, Europe, and North America, the most common food antigens causing CAFR are beef, dairy products, chicken, wheat, and lamb . In cats, the most common food antigens that are incriminated are beef, fish, and chicken . The best diagnostic procedure for identifying CAFR in companion animals is an elimination diet with subsequent provocation trials . To reliably diagnose CAFR in more than 90% of dogs and cats, elimination diet trials should last at least 8 wk and can be either homemade or commercially produced . Although veterinary dermatologists often consider home cooked diets as their first choice, many pet owners prefer the convenience of commercial “novel” or “hydrolyzed” protein diets, and rely on package labels to select diets that do not contain previously fed ingredients . There are concerns that these diets may contain unlisted food sources, and that unidentified ingredients might cause clinical reactions in hypersensitive patients . Unidentified dietary allergens could preclude a resolution of clinical signs in CAFR-affected patients, yielding misleading results for the elimination diet trial. Inadvertent cross-contamination of pet foods appears common, even in those with “limited ingredients” proposed for elimination diets . Rigorous quality control to screen for accidental contamination using real-time or quantitative polymerase chain reaction has been used in both human and animal food manufacturing as a rapid and sensitive point-of-care application to screen for food-borne bacteria, viruses, or allergens, square pots plastic which can be identified at low concentrations . This screening method has also been used in the production of veterinary prescription diets to validate their contents in order to avoid inadvertent allergen exposures for sensitized pets .
Of the quality control analysis methods available, DNA-based protocols are considered most reliable for detecting animal species in processed pet food manufacturing . Historically, these diets were fed to racing greyhounds and sled dogs; this feeding practice began to be extended to pets and became increasingly popular in the 1990’s . Despite evidence of nutritional deficiencies or excesses and potential health risks of feeding raw or undercooked animal source proteins, advocates of RMBD claim anecdotal health benefits, such as improvement in coat and skin, and a reduction in medical conditions such as allergies . The increasing popularity of RMBD has been demonstrated in multiple studies. A 2008 telephone survey revealed approximately 30% of dogs and 15% of cats in the United States and Australia consumed a combination of “unconventional” diets and commercial pet foods . A more recent anonymous Internet-based American survey indicated that 46% of dog owners and 38% of cat owners had fed RMBDs to their pets . Additionally, sales of RMBD have increased annually by as much as 15% in recent years . Over the past decade, this feeding practice has continued to increase, and market locations have expanded to include grocery stores, mass merchandisers, pet specialty stores, and veterinary clinics . A 2019 Italian-based survey shed light on dog owners’ motivations for adopting this feeding method . About 80% of respondents reported that they abandoned feeding commercial diets due to distrust in the clarity of ingredients commercial prescription veterinary diet, veterinarians may acquiesce to such requests. However, the reliability of RMBD for this purpose has not been evaluated. The primary aim of this study was to use PCR to test commercially available RMBD for the presence of DNA of animal origin other than that declared on the labels. A secondary objective was to determine the consistency of DNA presence between different batches of the same diets. The hypothesis was that the diets would contain unlisted protein ingredients, and that these unlisted proteins would vary between batches. To the authors’ knowledge, no previous studies have examined these issues.Contamination of one or both batches in all canine RMBD and most of the feline RMBD tested was detected in this study, which supports the hypothesis that cross-contamination would be found in many RMBD. Numerous independent studies have also demonstrated significant discrepancies between label claims and actual contents of dry or canned over-the-counter commercial diets, including those marketed for the management of CAFR . While this finding may have been expected in RMBD due to the prevalence of unlisted DNA detected in other such studies, diet purity could theoretically have been improved in RMBD as they are purported to undergo less processing before distribution, allowing less opportunity for protein contamination. An additional finding of this study was that unlisted animal source proteins varied among batches in most batches tested, including both canine and feline RMBD that were analyzed. Discrepancy among batches has not been previously studied for comparison, but these results showed that differences in unlisted ingredients were common in RMBD. Due to cost limitations restricting the analysis to only 2 batches of each RMBD, a statistically significant batch contamination rate could not be determined. However, the finding of discrepancy among batches represents yet another variable which could impact interpretation of an ED trial in a patient fed a commercially prepared RMBD. While no particular manufacturer’s diets were found to be more likely to contain unlisted proteins, brand F, the producer of feline diets 7 to 9, had the least number of contaminants, as well as the most consistent agreement between each batch. It is possible that this finding is due to the nature of the processing practices of this particular manufacturer, the production of smaller batch sizes to minimize opportunity for contamination, or more limited sourcing of ingredients to restrict the potential for supplier cross-contamination. Overall, the number of animal protein ingredients included in each diet was not a predictor of the number of unlisted proteins isolated in the analysis. Even diets restricted to single proteins were as likely to contain 1 or more sources of unlisted animal DNA as those with multiple animal proteins and batch contamination was unpredictable. Previous studies have shown that in rare cases, ingredients listed on product packaging were found to be missing from the analysis . Our study showed that no animal DNA was missing from that declared on the packaging of any RMBD included in the analysis. Due to the target DNA of the qPCR assay, the study was unable to validate the presence of sardine or goose to confirm the inclusion of these ingredients. This was again due to cost limitations precluding the addition of these proteins in the analysis. Some diets also contained animal fat sources such as salmon oil, cod oil, or sardine oil. Through purification processes, fish oils undergo refinement to remove proteins from the oil to render them free of proteins . While cod and sardine were not included in the analysis, our study did evaluate for salmon DNA. Both canine diets 1 and 3 contained salmon oil and tested negative for salmon DNA.