Forthe Presence Research Articles

Search for antibiotic-resistant strains of Gram-positive and Gram-negative microorganisms in poultry products

Research on the search for antibiotic-resistant strains of Gram-positive and Gram-negative microorganisms inpoultry products is an urgent and important topic in modern medicine and the food industry. Antibiotics arewidely used in poultry farming for the prevention and treatment of infections, but this also contributes to thedevelopment of antibiotic resistance in microorganisms. The purpose of this study is to identify and assess thelevel of antibiotic resistance of Gram-positive and Gram-negative bacteria present in meat and poultryderivedproducts. The methods include analyzing food samples, namely chicken fillets and chicken eggs, forthe presence and identification of resistant strains, as well as testing their sensitivity to various antibiotics.During the study, it was found that 19.05 % of Gram-positive strains and 26.09 % of Gram-negative strainsisolated from farm products exhibit resistance or multi-resistance properties. At the same time, 58.28 % ofGram-positive and 59.42 % of Gram-negative microorganisms also exhibit resistance or multi-resistanceproperties.

An update on apple chlorotic leaf spot virus status of sweet cherry in Bulgaria

The sweet cherry (Prunus avium L.) is one of the most important stone fruit species in Bulgaria. The cherry is susceptible to many viruses. To gain a better insight into the phyto-virologic status of sweet cherry, a survey for the presence of apple chlorotic leaf spot virus (ACLSV), cherry leafroll virus (CLRVR) and raspberry ringspot virus (RpRSV), in addition to ilarviruses, was carried out in 32 commercial and 6 collection orchards in ten regions of Bulgaria between 2017 and 2022. A total of 1503 sweet cherry samples were collected from symptomatic and symptomless trees. These samples were tested by cocktail ELISA for ACLSV and DAS-ELISA for CLRVR, RpRSV ?nd additionally for prune dwarf virus (PDV) and prunus necrotic ringspot virus (PNRSV) to detect mixed infections. Of the three viruses studied, only ACLSV was detected in commercial and collection sweet cherry orchards in all regions with intensive cherry cultivation in Bulgaria, in addition to ilarviruses. The extent of ACLSV virus infection was 8.8%. Single ACLSV infection was the most common, affecting 79.6% of infected sweet cherry trees, followed by ACLSV and PDV (14.4%) and least frequently by ACLSV and PNRSV (6.0%). The presence of ACLSV in orchards of different ages, including ?young? orchards, indicates that virus-free propagation material must be used when establishing new orchards to prevent the spread of pathogens with the planting material.

Molecular evidence of Theileria orientalis infection in cattle from Bosnia and Herzegovina

There are no data on the distribution of oriental theileriosis in cattle from Bosnia and Herzegovina. For the first time, a possible endemic focus of Theileria orientalis infection was confirmed in specific areas of Pale municipality, Sarajevo-Romanija region, Bosnia and Herzegovina. Selective sampling included 30 cattle from 10 smallholder farms in several locations in Pale municipality. The total of 30 whole blood samples were screened for the presence of piroplasms using commercial PCR. Positive PCR products were sequenced in both directions, the sequences were analyzed and a phylogenetic tree was created. Piroplasm (Babesia/Theileria-specific) DNA fragments were detected in 13/30 examined cattle (43%). At the farm level, PCR-positive animals were identified in 6/10 examined farms (60%). Upon sequence analysis, the species Theileria orientalis was confirmed. This survey reports a high rate of PCR-positive cases of bovine piroplasmosis and provides the first description of Theileria orientalis in Bosnia and Herzegovina.

ANTIPLASMODIAL ACTIVITY AND PHYTOCHEMICAL ANALYSIS OF EXTRACTS FROM THE ROOTS AND STEM BARK OF JATROPHA CURCAS

The powdered roots and stem bark of Jatropha curcas were extracted using 95% ethanol. The ethanol extracts obtained from parts of plants were partitioned into fractions using chloroform, distilled water, ethyl acetate, methanol and n-hexane solvents of varying polarity. The crude ethanol extract and five soluble fractions of both the roots and stem bark were tested against plasmodium falciparum. The results shown a good anti-plasmodial potency at high concentration of the fraction of the roots and stem bark of J. curcas. Chloroform and n-hexane soluble fractions recorded good activity on the test organisms with 94.3% and 97.1% inhibition. The results indicated that the roots and stem bark extracts contained active compounds which have anti-malaria property. Test for the presence of secondary metabolites in the plant extracts revealed the presence of saponins, flavonoids, steroids, tannins and alkaloids.

Prevalence of Upper Quadrant Musculoskeletal Disorders among Healthcare Professionals in a Tertiary Care Hospital

Background: Healthcare professionals (HCPs) are vulnerable to acquire musculoskeletal disorders during theirwork routine. Shortage of HCPs, long working hours and mandatory overtime results in overburden on thoseavailable, especially during pandemic such as in COVID -19. Objective of the study was to evaluate the prevalenceof upper quadrant musculoskeletal disorders (UQMSDs) among HCPs.Methodology: A total of 370 HCPs including doctors, dentists, physiotherapists and nurses were evaluated forthe presence of trigger points (TrPs) in upper trapezius, levator scapula, infraspinatus and sternocleidomastoidmuscles, severity of pain (NPRS), neck disability (NDI) and cervical and shoulder ROM (goniometer).Results: The prevalence rate of UQMSDs was found to be high among HCPs. TrPs were more prevalent in uppertrapezius muscle (46.21%) followed by levator scapulae, infraspinatus and sternocleidomastoid muscles. 4.1% ofHCPs reported severe pain, 16.7% reported moderate pain and 48.36% reported mild pain in upper quadrant.0.8% of HCPs had complete, 1.2% had severe, 5.3% had moderate and 37.6% had mild neck disability. The cervicalrotation and shoulder horizontal adduction ROM were found to be more restricted among HCPs.

Assessment of Children with Thumb Sucking Habit in Babylon City

Aims: The purpose of this study is to assess the prevalence of thumb sucking habit between preschoolersand the effect on their occlusion of the habit.Materials and Methods: 42 of pre-school children aged 4-5 years who were inspected by questioning forthe presence or absence of thumb sucking habit, and thumb examination, and the teeth were examined forthe presence of decreased over flight and overbite or open bite.Results: Total number children have thumb sucking habit, and open bite of them.Conclusion: In this age group (4-5) years , the number of children with thumb sucking habit was consideredhigh and the habit was found to increase the prevalence of malocclusion of deciduous.

Effect of Aqueous Warionia saharae Extract on Lipid and Glucose Metabolism in Normal and Diabetic Rats

Objective:The aim of the study was to evaluate the hypolipidemic and antioxidant properties of aerial parts aqueous extract of Warionia saharae (APAE of W. saharae) in normal and streptozotocin (STZ) induced-diabetic rats. Additionally, the quantitative and qualitative analysis for the presence of different phytochemical constituents in the APAE of W. saharae was performed.Methods:The effects of oral administration of APAE of W. saharae (5 mg/kg) on the plasma Total Cholesterol (TC) and triglyceride (TG) levels were measured in both normal and diabetic rats. Moreover, the antioxidant capacity was realized by the method of DPPH. Total polyphenol as x of the APAE of W. saharae were determined.Results:APAE of W. saharae showed a strong hypolipidemic and antihyperglycemic effects both in normal and in STZ induced diabetic rats. On the other hand, APAE of W. saharae showed a potent antioxidant activity and revealed inhibitory activity. Moreover, the quantitative determination of total polyphenol and flavonoid contents showed high contents of these phytochemicals. Concerning the qualitative analysis several classes of chemicals have been found.Conclusion:APAE of W. saharae (5 mg/kg) shows the potential cholesterol and glucose lowering activity as well as antioxidant properties in both normal and STZ-induced diabetic rats.

Phytochemical Investigation and Assessment of Antioxidant and Antimicrobial Activities of Phagnalon barbeyanum Aerial Parts

Objectives: The present study aimed to investigate and isolate different phytochemical constituents and assess the antioxidant and antimicrobial activities of the total methanol extract and different fractions of Phagnalon barbeyanum aerial parts. Methods: Phytochemical screening of Phagnalon barbeyanum aerial parts was performed using specific test for each class of compounds. Different chromatographic techniques were used to isolate and purify compounds and their structures were elucidated by using different spectral techniques (1H NMR and 13C NMR). The antioxidant activity was evaluated by DPPH radical scavenging assay and antimicrobial activity was done by standard agar well diffusion assay. Results: Phytochemical investigation of Phagnalon barbeyanum aerial partswas done forthe presence ofcarbohydrates and or glycosides, sterols and or triterpene, flavonoids, tannins and saponins and revealed the absence of alkaloids and anthraquinones. Compounds; β-sitosterol (1), apigenin (2) and β-sitosterol-3-O-β-D- glucopyranoside (3) were isolated from methylene chloride and ethyl acetate fractions. The ethyl acetate fraction showed highest antioxidant activity followed by n-butanol and methylene chloride fractions. The total methanol extract and n-hexane fractions showed lowest antioxidant activity. The ethyl acetate fraction and total methanol extract showed moderate activity against Staphylococcus aureus and Vancomycin resistant Staphylococcus aureus. Conclusion: The aerial parts of Phagnalon barbeyanum is a rich source of different classes of active constituents as phenolics. The total methanol extract of Phagnalon barbeyanum aerial parts and some of its fractionated concentrates could be considered as antioxidant and antimicrobial agents.

Nutraceuticals from Marine Bionetworks

“Nutrition” and “Pharmaceutical” together build up the perception of “Nutraceuticals” that refer to the food or dietary supplements that help to incorporate additional health benefits to the fundamental sustenance accomplished on daily basis. Each nutraceutical contains one or more bioactive molecules that are usually obtained by chemical and/ or biotechnological synthesis or by extraction from natural sources. Among the natural sources, marine bionetwork possess immense potential for the presence of bioactive compounds. Some of these bioactive compounds as isolated from marine sources, have potential use as nutraceuticals. This mini review provides a brief overview of nutraceutical compounds from marine sources that are currently under research and/or have been commercialized. A detailed discussion on the biochemical categories of compounds and the marine organisms that play as potential sources of these bioactive nutraceutical compounds have been included.

Occurrence of neonatal diarrhea in calves with iron-deficiency anemia

Introduction. Neonatal calves are often deficient in iron. Accumulating evidence indicates that iron status is associated with disease pathologies including diarrhea. Our objective was to examine the association between iron status and gut function in neonatal calves with and without a history of calf diarrhea. Materials and Methods. Calves were divided into two groups based on their history of diarrhea; the first group were diarrheic calves (n=6) and the second group were non-diarrheic healthy calves (n=6). Blood samples (n=12) were collected at day 12 of age and erythrogram determination and measurements of serum iron and total iron binding capacity were performed. Hematological values were measured using an automatic analyzer, and biochemical properties were determined spectrophotometrically. Fecal samples were obtained from all calves and pH measured using semi quantitative test strips as well as being examined by bacterial cultivation for enterotoxigenic Escherichia coli, Salmonella spp. and Clostridium perfringens, by RT-PCR for the presence of bovine rotavirus, bovine coronavirus and bovine viral diarrhea virus, and by microscopy for the presence of Cryptosporidium parvum. Results and Conclusions. There were significant iron-related changes for most hematological indices in diarrheic calves; and iron (Fe) deficiency and microcytic, hypochromic anemia were diagnosed. The pH of the feces was significantly higher in diarrheic calves than in the non-diarrheic healthy group (P<0.01). All fecal samples were negative for the analyzed enteric pathogens. According to the results obtained, calves experiencing iron deficiency anemia exhibit changes in gut function leading to diarrhea as compared with a matched group of healthy calves.

Southernmost records of Escarpia spicata and Lamellibrachia barhami (Annelida: Siboglinidae) confirmed with DNA obtained from dried tubes collected from undiscovered reducing environments in northern Chile.

Deep-sea fishing bycatch enables collection of samples of rare species that are not easily accessible, for research purposes. However, these specimens are often degraded, losing diagnostic morphological characteristics. Several tubes of vestimentiferans, conspicuous annelids endemic to chemosynthetic environments, were obtained from a single batch of deep-sea fishing bycatch at depths of around 1,500 m off Huasco, northern Chile, as part of an ongoing study examining bycatch species. DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene and an intron region within the hemoglobin subunit B2 (hbB2i) were successfully determined using vestimentiferans’ dried-up tubes and their degraded inner tissue. Molecular phylogenetic analyses based on DNA sequence identified the samples as Escarpia spicata Jones, 1985, and Lamellibrachia barhami Webb, 1969. These are the southernmost records, vastly extending the geographical ranges of both species from Santa Catalina Island, California to northern Chile for E. spicata (over 8,000 km), and from Vancouver Island Margin to northern Chile for L. barhami (over 10,000 km). We also determined a 16S rRNA sequence of symbiotic bacteria of L. barhami. The sequence of the bacteria is the same as that of E. laminata, Lamellibrachia sp. 1, and Lamellibrachia sp.2 known from the Gulf of Mexico. The present study provides sound evidence forthe presence of reducing environments along the continental margin of northern Chile.

On nature’s shoulders

The use of counterfactual methods in the evaluation of policy interventions has been accepted today as the best approach in the estimation of a program’s performance. However, the simplest evaluations are often quite demanding in terms of the resources and the time needed to be implemented. In this article, we study the economic impact of a tourism media campaign launched in Nazare, an old fishing community on the west coast of Portugal, to make big waves visible to the world. The campaign provided the required “informational media infrastructure” that created the public awareness necessary to boost tourism in the region. To measure the economic impact of that campaign on the local economy, we show how a counterfactual analysis can be implemented using regional statistical data on domestic and international tourist arrivals. We show how the method can be adapted to account for the presence of potential spillover effects that may have occurred, as neighboring municipalities could also have been affected by the intervention. We further compare the estimated impact on revenues with the costs incurred by the local municipality in the marketing campaign. Based on our empirical findings, we discuss policy implications to the municipalities in the region.

A STUDY ON STANDARDIZATION AND ASSESSMENT OF SENSORY AND MICROBIOLOGICAL QUALITY OF HERBAL RASAM

Objective: Herbs have gained a widespread acceptance as a dietary constituent in Tamil Nadu, generally forming a substantial portion of the dietwhich serves as a therapeutic function. This study was aimed to formulate and standardize the four types of Rasam using four different herbs and toanalyze its quality changes in fresh Rasam and refrigerated Rasam (4±1°C). Appearance and odor scores of Rasam were satisfactory throughout thestorage. Sensory attributes were rated better for fresh Rasam than refrigerated Rasam, but all the attributes were decreased with increasing storageperiod.Methods: Microbiological quality of four types of different herbal Rasam was examined with freshly prepared Rasam and with refrigerated Rasam forthe presence of Escherichia coli, Staphylococcus aureus, Salmonella and Yeast, and Mold.Result: Comparison with the microbiological results shows in refrigerated Rasam-PAR, SAR, and AIR were at an acceptable level of microbiologicalquality whereas, Rasam-STR were at an unacceptable level of microbial quality due high level of total plate count and yeast and mold growth.Salmonella was examined, but it was not detected in all the four Rasam. Similarly, the microbial quality of all four freshly prepared Rasam showedhighly acceptable. Conductivity was measured for the presence of ions in solution.Conclusion: This study proposes that to minimize bacterial level in foods and always to consume freshly prepared Rasam than refrigerated. It alsoconcludes that good hygiene practices are necessary though the food serves a therapeutic function.

ISOLAMENTO E IDENTIFICAÇÃO DE FUNGOS DO GÊNERO Aspergillus spp. DE ÁGUA UTILIZADA NA REABILITAÇÃO DE PINGUINS-DEMAGALHÃES

Resumo A aspergilose caracteriza-se por ser a principal causa de mortalidade de pinguins em cativeiro. A infecção pelo gênero Aspergillus ocorre principalmente por via aérea, porém o fungo pode ter dispersão pela água. Neste sentido, este trabalho teve como objetivo avaliar a água do tanque onde os pinguins-de-Magalhães permanecem para reabilitação no Centro de Recuperação de Animais Marinhos de Rio Grande, RS, Brasil, quanto à presença de fungos filamentosos do gênero Aspergillus. As amostras de água foram coletadas semanalmente durante um período de 10 meses e processadas em um período máximo de seis horas utilizando-se a técnica da membrana filtrante, com incubação a 25 ºC e 37 ºC por até sete dias. Das 40 amostras analisadas, 32 foram positivas para o isolamento do gênero Aspergillus, sendo que dessas 60% pertenciam à espécie A. fumigatus. Algumas variáveis interferiram significativamente no isolamento do gênero Aspergillus e/ou da espécie A. fumigatus, como temperatura de incubação, sazonalidade e densidade populacional. Este trabalho demonstra que Aspergillus spp. está presente na água, podendo essa ser uma potencial fonte de infecção para os pinguins em reabilitação.

Risk factors for intraocular involvement in patients with primary central nervous system lymphoma.

To determine the risk factors for intraocular involvement in patients with primary central nervous system lymphoma (PCNSL), a retrospective chart review was performed on 136 patients who were pathologically diagnosed with PCNSL. The patients were investigated for demographics, clinical manifestation, and the profile of immunohistochemical tumor biomarkers, as well as forthe presence of intraocular involvement of lymphoma at diagnosis or during follow-up. The mean age of the entire cohort was 58.6±12.4years, and the mean follow-up period was 31.1±30.8months. Twenty-nine (21%) patients had an intraocular involvement, among which 20 (69%) patients presented with intraocular involvement at diagnosisof PCNSL and 9 (31%) patients developed intraocular involvement after a mean period of 32.4±33.6months. Of the patients with intraocular involvement, 8 (28%) had no visual symptom at the diagnosis of ocular invasion. Between those with and without intraocular involvement, no significant differences were found with respect to the age, sex, and follow-up period as well as cerebrospinal fluid spread and bone marrow involvement. Among the immunohistochemical biomarkers, the Ki-67 proliferation index was significantly higher in patients with intraocular involvement than in patients without (P=0.021), but the other investigated biomarkers did not show a significant difference between the two groups. A Ki-67 level ≥80% was a risk factor for the intraocular involvement in patients with PCNSL (odds ratio, 2.63). Median overall survival was 39.0months in the entire cohort and was not significantly different between those with and without intraocular involvement (P=0.959).

Quantitative and Pattern Analyses of Continuous-Wave Doppler–Derived Pulmonary Regurgitant Flow Velocity for the Diagnosis of Constrictive Pericarditis

Many echocardiographic features of constrictive pericarditis (CP) have been reported, but each alone has a limitation either in sensitivity or in specificity. Continuous-wave Doppler-derived flow velocity of pulmonary regurgitation can reflect the diastolic right ventricular pressure pattern characteristic of CP and be useful for its detection. Fifteen patients with CP, 18 patients with restrictive cardiomyopathy, and 20 normal subjects were studied retrospectively. Using continuous-wave Doppler echocardiography, pulmonary regurgitation velocities were measured at the early diastolic peak (VMAx), mid-diastolic inflection point (VIFL), and late diastolic minimal point (VMIN). VIFL, VIFL/VMAx, VMIN, and VMIN/VMAx were significantly lower in the CP group compared with the restrictive cardiomyopathy and normal groups. Prevalence rates of patients with early mid-diastolic inflection, VIFL/VMAx < 0.5, VMIN < 50 cm/sec, and VMIN/VMAx < 0.33 were significantly greater in the CP group compared with the other groups. Sensitivity and specificity for the diagnosis of CP were 93% and 74%, respectively, forthe presence of early mid-diastolic inflection, 73% and 100% for VIFL/VMAx < 0.5, 73% and 97% for VMIN < 50 cm/sec, and 93% and 92% for VMIN/VMAx < 0.33. The quantitative and pattern analyses of continuous-wave Doppler-derived pulmonary regurgitation velocity could enhance the accuracy of echocardiographic detection of CP.

Presence of Illicit Drugs in the Sarno River (Campania Region, Italy)

The presence of illicit drugs and their metabolites in surface waters has to be considered a new type of hazard, still unknown, for both the human health and the aquatic ecosystem, due to the potent pharmacological activities of all the illicit drugs. Our research was aimed at evaluating the presence of illicit drugs in the Sarno River (Campania region, Italy), crossing a densely populated area, the basin of the Sarno River, one of the largest and most important economic areas in Campania region, famous for the presence of zones with high landscape-environmental value. The drugs selected for this study were the most used in Campania region. The presence of illicit drugs in surface water was analyzed by a selective multi-residue assay based on liquid chromatographytandem mass spectrometry. The analysis showed the presence of all the illicit drugs investigated: cocaine and its main metabolites (benzoylecgonine, nor-benzoylecgonine), morphine, THC-COOH and codeine; cocaine was the most abundant illicit drug. The presence of illicit drugs and their metabolites in the Sarno River suggests new potential risk for the basin’s inhabitants, using water in the food chain, via field irrigation and animal feed, and for the health of the aquatic fauna.

Spatial Distribution of Plant-Associated Invertebrates and Environmental Bioassessment in a Natural Riverine Lake (Lake Fibreno, Central Italy)

Lake Fibreno is a Mediterranean riverine lake located in Central Italy and fed by large flows of groundwater and characterized by the presence of a wide reed stand area and of the only floating island of the South Europe. It is enclosed in a Natural Reserve recognized as Special Area of Conservation (SAC-IT6050015) for the presence of habitats and species (92/43/EEC) of relevant scientific interest. In this paper, we compare the plant-associated meio-macrobenthic invertebrates among sites with different environmental characteristics. Both vegetation and invertebrates of Lake Fibreno were well diversified and abundant and were composed of a mixing of species typical of lentic and lotic waters. Our results show that a gradient of orthophosphate and nitrite contents and current velocity appeared to be the driver factors affecting the flora and fauna spatial distribution at the sampling sites. To obtain a bioassessment of the ecological quality of the vegetation zone, we compared the riverine Lake Fibreno with typical lentic lakes previously studied in Central Italy. We selected metrics, such as macrophyte belt extent and taxa richness, invertebrate species richness and Shannon diversity, ETOM (Ephemeropta + Trichoptera + Odonata + Mollusca), ETOC (Ephemeropta + Trichoptera + Odonata + Coleoptera) and LCrM (Large Crustacea + Mollusca). The comparison indicated a good ecological status of the vegetation zone of Lake Fibreno mainly due to the continuous washout effects (dilution and fast water removal) which reduce the nutrient content available for phytoplankton and maintain the high transparency and good oxygenation of the lake waters.

Monoclonal antibody technology applied to the detection of &amp;lt;i&amp;gt;Cryptosporidium parvum&amp;lt;/i&amp;gt; oocysts in human and cattle fecal samples

With the discovery that the coccidian parasite Cryptosporidium sp. can cause severe symptoms in humans, many diagnostic techniques were quickly implemented such as microscopic visualization, immunofluorescence and PCR. Currently, there is no effective drug treatment and none of the current diagnostic methods is 100% accurate. In this study, a BALB/C mouse was subcutaneously immunized with Cryptosporidium parvum oocysts extract. The spleen was removed and the splenocytes were fused with SP2/0 myeloma cells in order to obtain hybridoma cells secreting antibodies specific to C. parvum antigens. Human and cattle fecal samples previously characterized by microscopy [Ziehl-Neelsen staining (ZN) and Lugol] and PCR for the presence of C. parvum and Giardia duodenalis, were analyzed by indirect immunofluorescence, using the developed hybridomas supernatants. The study shows that the selected hybridomas supernatants identify C. parvum oocysts in fecal samples in correlation with C. parvum oocysts identified using ZN/PCR. No false positive results were obtained and the two best supernatants gave 20% -30% of false negative results. No cross reaction with G. duodenalis was observed. By comparing our results with those obtained with an immunofluorescence commercial kit, it suggests the potential use of the monoclonal antibodies present in two of the hybridomas supernatants as a detection tool of C. parvum. With a reliability of 80.8% and 73.1% versus ZN and PCR methods for IFI, compared with a reliability of 76.9% and 92.3% versus ZN and PCR for commercial DIF kit, the supernatant 4.1D5 seems to be the most promising subject to further study its usefulness for C. parvum detection.

Illicit Drug Use in the Pain Patient Population Decreases with Continued Drug Testing

Background: A major concern of physicians treating pain patients with chronic opioid therapy and similar drugs is determining whether the patients are also using illicit drugs. This is commonly determined by urine drug testing (UDT). However, there are few studies on whether or not monitoring patients by this technique decreases illicit drug use. Objective: To determine if the presence of illicit drugs decreases over a number of physician visits where UDT was performed. Method: The method involved a retrospective study of tests for the illicit drugs marijuana, cocaine, methamphetamine, ecstacy (MDMA) phencyclidine (PCP) and the heroin metabolite, 6-acetylmorphine as confirmed by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). A database of 150,000 patient visits was examined for the presence of any of these 6 drugs. Results: A total of 87,000 patients were initially tested. The number of patients who were repeatedly tested decreased over time. The percentage of patients positive for any of these illicit drugs decreased from 23% to 9% after 14 visits where UDT was performed. When graphed there was a trend to decreasing use. The Spearman correlation = -0.88, P &lt; 0.0001. The major illicit drug was marijuana. When this was removed from the analysis, there was an even greater correlation with decreased illicit drug use. Spearman correlation = -0.92 (P &lt; 0.0001) using a weighted correlation. Limitation: Patients continuing to use illicit drugs might be dismissed from practices thus biasing the study towards illicit drug avoidance. Conclusion: Continued UDT might decrease illicit drug use among pain patients. Key words: Pain patients, UDT, urine drug testing, LC-MS/MS, illicit drugs, decrease drug use