Dietary inflammatory index and mortality: a cohort longitudinal study in a Mediterranean area.

BACKGROUND: Higher Dietary Inflammatory Index (DII®) scores are associated with increased morbidity and mortality. However, little is known about the effects of DII on mortality in Mediterranean countries. Therefore, in the present study, we aimed to investigate the potential association between DII scores and overall, cancer and cardiovascular disease (CVD) mortality in people living in a Mediterranean area. METHODS: DII scores were calculated using a validated food-frequency questionnaire. DII scores were then categorised into tertiles. Mortality was ascertained via death certificates. The association between DII scores with overall and cause-specific mortality was assessed via a multivariable Cox's regression analysis and reported as hazard ratios (HRs) with their 95% confidence intervals (CIs). RESULTS: The study included 1565 participants (mean age 65.5 years; females 44.7%). After a median follow-up of 12 years (2005-2017), 366 (23.4%) participants died. After adjusting for 17 potential confounders, people with higher DII scores had an increased risk of death compared to those in the lowest (most anti-inflammatory) tertile (HR = 1.38; 95% CI = 1.04-1.82 for the second tertile; HR = 1.38; 95% CI = 1.03-1.86 for the third tertile). Each 1 SD increase in DII score increased the risk of death by 13%. No association was found between DII scores and cancer or CVD death when considered separately. CONCLUSIONS: Higher DII scores were associated with a significantly higher mortality risk, whereas the association with cause-specific mortality was less clear. These findings highlight the potential importance of diet in modulating inflammation and preventing death.

Functional foods and cardiometabolic diseases* International Task Force for Prevention of Cardiometabolic Diseases.

Mounting evidence supports the hypothesis that functional foods containing physiologically-active components may be healthful. Longitudinal cohort studies have shown that some food classes and dietary patterns are beneficial in primary prevention, and this has led to the identification of putative functional foods. This field, however, is at its very beginning, and additional research is necessary to substantiate the potential health benefit of foods for which the diet-health relationships are not yet scientifically validated. It appears essential, however, that before health claims are made for particular foods, in vivo randomized, double-blind, placebo controlled trials of clinical end-points are necessary to establish clinical efficacy. Since there is need for research work aimed at devising personalized diet based on genetic make-up, it seems more than reasonable the latter be modeled, at present, on the Mediterranean diet, given the large body of evidence of its healthful effects. The Mediterranean diet is a nutritional model whose origins go back to the traditional dietadopted in European countries bordering the Mediterranean sea, namely central and southern Italy, Greece and Spain; these populations have a lower incidence of cardiovascular diseases than the North American ones, whose diet is characterized by high intake of animal fat. The meeting in Naples and this document both aim to focus on the changes in time in these two different models of dietary habits and their fall out on public health.

Virus C hepatitis and type 2 diabetes: a cohort study in southern Italy.

OBJECTIVES: The relationship between hepatitis C virus (HCV) infection and type 2 diabetes mellitus (DM 2) is still uncertain. The objective of this study was to evaluate the association between HCV infection, measured as positivity to anti-HCV antibodies, and the incidence of DM 2 in a cohort of subjects sampled from the general population and followed up for 20 years. METHODS: At baseline, the cohort consisted of a random sample of 2,472 subjects (72% response rate, age range 30-69 years) from the electoral register of a town in Southern Italy. The cohort subjects were examined three times: in 1985 (M1), in 1992 (M2), and in 2005 (M3). At M1, M2, and M3, each participant filled in a questionnaire and had a blood sample taken to measure blood glucose and other serum variables including glutamic pyruvic alanine aminotransferase (ALT). Anti-HCV antibodies were analyzed with standard techniques at M1 and M2. Diabetes type 2 diagnosis was a history of diabetes and/or serum glucose ≥126 mg/dl and/or treatment with insulin or hypoglycemic drugs. Logistic regression was used for multivariable data analysis. RESULTS: Diabetes prevalence was higher in subjects with positive anti-HCV antibodies at M1 and M2, and diabetes incidence was higher in subjects with baseline positive anti-HCV antibodies at M1-M2 and lower at M2-M3. In multivariable models, controlling for gender, age, and body mass index (BMI), there was no association between incident cases of diabetes and positive anti-HCV antibodies at baseline, either at M1-M2 (odds ratio (OR) 0.73, 95% confidence interval (CI) 0.43-1.22) or at M2-M3 (0.65, 0.41-1.04). HCV was associated with DM 2 only in subjects with elevated ALT (OR 0.58, 95% CI 0.31-1.08, if ALT normal; OR 1.47, 95% CI 1-2.16, if ALT elevated, controlling for age, gender, and BMI). CONCLUSIONS: Our findings, in a cohort study at population level, support an association between the presence of anti-HCV antibodies at baseline and a higher incidence of type 2 diabetes in the following 20 years only in subjects with elevated ALT.

Diet and myocardial infarction: a nested case-control study in a cohort of elderly subjects in a Mediterranean area of southern Italy.

BACKGROUND AND AIM: We evaluated the incidence of myocardial infarction (MI) in a population of Southern Italy and the relationship of dietary macronutrients with incident MI. METHODS AND RESULTS: The ONCONUT cohort included 5632 subjects followed-up, over 50 years, recruited in 1992. At baseline, they completed a validated semi-quantitative food frequency questionnaire and gave details of their medical history. After 5years they were traced by their family physician, who found 108 incident MI. Ninety-seven of them and 194 controls, sampled from the noncases at baseline and paired for diabetes to the cases, entered this nested case-control study. MI rate per 1000 person-years was 9.6 in males and 3.7 in females. In non-diabetics, saturated fat were associated with MI directly (odds ratio (OR): tertile 2 vs. 1 = 2.32, tertile 3 vs. 1 = 2.82; chi-square for trend, p = 0.03) and polyunsaturated fats inversely (OR: tertile 2 vs. 1 = 0.80, tertile 3 vs. 1 = 0.37; chi-square for trend, p = 0.05), while in diabetics, starchy carbohydrates (OR: tertile 2 vs. 1 = 1.51, tertile 3 vs. 1 = 6.73; chi-square for trend, p = 0.01) and glycaemic index (OR: tertile 2 vs. 1 = 2.74, tertile 3 vs. 1 = 5.34; chi-square for trend, p = 0.01) were associated directly with MI. CONCLUSIONS: MI incidence in this population was lower than that found in northern countries. In non-diabetics, saturated fats were associated directly and polyunsaturated fat inversely with MI; in diabetics, starchy carbohydrates and high-glycaemic-index foods were associated directly with MI.

Fatty liver and mortality: a cohort population study in South Italy.

OBJECTIVE: Alcoholic fatty liver (AFLD) and non-alcoholic fatty liver (NAFLD) are two common conditions. However, if they can increase the risk of death is poorly explored. We therefore aimed to investigate the potential association between the presence and severity of liver steatosis and mortality in a large sample of older people. DESIGN: Prospective. SETTING: Community. PARTICIPANTS: Women and men randomly sampled from the electoral rolls of the population of Castellana Grotte, a town in Southern Italy (Apulia region) between 2005 and 2006. Among 1942 initially contacted, 1708 (=87.9%) participated to the baseline survey (Multicentrica Colelitiasi III (MICOL III)). This specific study included 1445 older participants (mean age=65.2 years, females=44.2%). EXPOSURE: NAFLD or AFLD. PRIMARY AND SECONDARY OUTCOMES: Mortality (all-cause and specific-cause). RESULTS: After a median of 12 years, 312 participants (=21.6%) died. After adjusting for nine potential confounders, the presence of steatosis was not associated with any increased risk of death in both NAFLD and AFLD. The severity of liver steatosis was not associated with any increased risk of mortality in NAFLD, while in AFLD, the presence of moderate steatosis significantly increased the risk of overall (HR=2.16; 95% CI 1.19 to 3.91) and cancer-specific (HR=3.54; 95% CI 1.16 to 10.87) death. CONCLUSIONS: Liver steatosis is not associated with any increased risk of death in NAFLD, while moderate steatosis could be a risk factor for mortality (particularly due to cancer) in people affected by AFLD.

Potato Consumption Is not Associated with Higher Risk of Mortality: A Longitudinal Study among Southern Italian Older Adults.

OBJECTIVE: The consumption of potatoes is increasing worldwide, but few studies have assessed the association between potato consumption and mortality, particularly in Mediterranean countries. We therefore investigated whether potato consumption is associated with higher risk of death in a large cohort of people living in South Italy. DESIGN: Longitudinal. SETTING: Community-dwelling. MEASUREMENTS: 2,442 participants coming from MICOL and NUTRIHEP studies aged more than 50 years at baseline were followed-up for 11 years. Dietary intake was assessed by means of a Food Frequency Questionnaire. Potato consumption was categorized in quintiles according to their daily consumption (< 3.95, 3.96-8.55, 8.56-15.67, 15.68-22.0, and > 22.0 g/day). Mortality was ascertained through validated cases of death. The association between potato consumption and mortality was assessed through Cox's regression models, adjusted for potential confounders, and reporting the data as hazard ratios (HRs) with 95% confidence intervals (CIs). RESULTS: The 2,442 eligible participants were prevalently males (54.6%) and aged a mean of 64.3±9.3 years. During the 11-year follow-up, 396 (=16.2%) participants died. After adjusting for 12 potential baseline confounders, and taking those with the lowest consumption of potatoes as the reference group, participants with the highest consumption of potatoes did not have an increased overall mortality risk (HR=0.75; 95%CI: 0.53-1.07). Modelling the potato consumption as continuous (i.e. as increase in 10 g/day) did not substantially change our findings (fully-adjusted HR=0.93; 95%CI: 0.84-1.02). CONCLUSION: Overall potato consumption was not associated with higher risk of death in older people living in a Mediterranean area. Future studies are warranted to elucidate the role of potato consumption on all-cause and cause-specific mortality.

Glycated apolipoprotein B and myocardial infarction.

AIMS: To evaluate the association of serum concentrations of glycated apolipoprotein B (ApoBg) with the incidence of myocardial infarction (MI) in subjects with and without diabetes. METHODS: The design is a nested case-control study. The cohort included 5632 subjects over 50 years of age attending the clinical laboratories of a small geographic area in southern Italy. After five years, 4563 subjects were traced and 103 had developed MI. We sampled from the cohort two controls for each incident case of MI, frequency matched for sex and diabetes. ApoBg was measured using a monoclonal antibody. Logistic regression was used for statistical analysis of the data. RESULTS: ApoBg at baseline was higher in subjects who developed myocardial infarction than in controls in both non-diabetic and diabetic subjects (t test, P=0.009 and P=0.05 respectively). MI odds ratio in the third tertile of ApoBg was 2.01 (95% CI 0.93-4.33) in non-diabetic and 2.88 (0.85-9.68) in diabetic subjects (chi-square test for trend; non-diabetics P=0.03, diabetics P=0.06). Serum triglycerides, cholesterol, HDL and LDL cholesterol, glucose and insulin were not associated with MI (P>0.10). CONCLUSION: ApoBg at baseline is directly associated with the development of MI in the following five years in both diabetic and non-diabetic individuals.

Effect of a Low Glycemic Index Mediterranean Diet on Non-Alcoholic Fatty Liver Disease. A Randomized Controlled Clinici Trial.

INTRODUCTION: Non-Alcoholic Fatty Liver Disease (NAFLD) is currently the most common form of liver disease worldwide affecting all ages and ethnic groups and it has become a consistent threat even in young people. Our aim was to estimate the effect of a Low Glycemic Index Mediterranean Diet (LGIMD) on the NAFLD score as measured by a Liver Ultrasonography (LUS). DESIGN: NUTRIzione in EPAtologia (NUTRIEPA) is a population-based Double-Blind RCT. Data were collected in 2011 and analyzed in 2013-14. SETTING/PARTICIPANTS: 98 men and women coming from Putignano (Puglia, Southern Italy) were drawn from a previous randomly sampled population-based study and identified as having moderate or severe NAFLD. INTERVENTION: The intervention strategy was the assignment of a LGIMD or a control diet. OUTCOME MEASURES: The main outcome measure was NAFLD score, defined by LUS. RESULTS: After randomization, 50 subjects were assigned to a LGIMD and 48 to a control diet. The study lasted six months and all participants were subject to monthly controls/checks. Adherence to the LGIMD as measured by Mediterranean Adequacy Index (MAI) showed a median of 10.1. A negative interaction between time and LGIMD on the NAFLD score (-4.14, 95% CI -6.78,-1.49) was observed, and became more evident at the sixth month (-4.43, 95%CI -7.15, -1.71). A positive effect of the interaction among LGIMD, time and age (Third month: 0.07, 95% CI 0.02, 0.12; Sixth month: 0.08, 95% CI 0.03,0.13) was also observed. CONCLUSIONS: LGIMD was found to decrease the NAFLD score in a relatively short time. Encouraging those subjects who do not seek medical attention but still have NAFLD to follow a LGIMD and other life-style interventions, may reduce the degree of severity of the disease. Dietary intervention of this kind, could also form the cornerstone of primary prevention of Type 2 Diabetes Mellitus (T2DM) and cardiovascular disease.

Diet, physical activity, and gallstones--a population-based, case-control study in southern Italy.

BACKGROUND: Gallstones are a highly prevalent condition; however, the nutritional and lifestyle risk factors of this disease are not well understood. OBJECTIVE: We evaluated the association between diet, physical activity, and incident cases of gallstones diagnosed by ultrasound in a population-based, case-control study. DESIGN: One hundred patients with newly diagnosed gallstones and 290 randomly selected control subjects without gallstones were enrolled in the study. The presence of gallstones was determined by ultrasonography. Both patients and control subjects completed a questionnaire about their usual diet and physical activity for the 12 mo before the ultrasonography. The association between diet and physical activity and risk of gallstone formation was analyzed by using multiple logistic regression. RESULTS: Body mass index and intake of refined sugars were directly associated with risk of gallstone formation, whereas physical activity, dietary monounsaturated fats, dietary cholesterol, and dietary fibers from cellulose were inversely associated with risk of gallstone formation. Saturated fats were a risk factor for gallstone formation and the association appeared to be stronger for men than for women. CONCLUSION: These findings suggest that a sedentary lifestyle and a diet rich in animal fats and refined sugars and poor in vegetable fats and fibers are significant risk factors for gallstone formation.

Markers of insulin resistance and colorectal cancer mortality.

A link between insulin metabolism and colorectal cancer has been hypothesized, supported by a series of potential physiological mechanisms, and confirmed by a number of reports in experimental animals. However, the evidence in humans is limited and mostly indirect. The aim of the present report is to analyze whether individuals with a cluster of metabolic abnormalities associated with abnormalities in insulin metabolism experience higher mortality for colorectal cancer than those without this cluster of metabolic abnormalities. A total of 21,311 men and 15,991 women 20-69 years of age were followed-up for an average of 7 years as part of the Risk Factors and Life Expectancy Project, a pooling of a number of epidemiological studies conducted in Italy. Our analyses indicate that participants with high levels of blood glucose and a cluster of metabolic abnormalities linked to insulin resistance experienced a significant increased risk of colorectal cancer mortality compared with participants without the cluster. For the presence of the cluster of metabolic abnormalities, the calculated hazard ratios and 95% CIs were 2.96 (1.05-8.31) for men, 2.71 (0.59-12.50) for women, and 2.99 (1.27-7.01) when both sexes were combined. These associations were independent from the potential confounding effect of age, drinking of alcoholic beverages, and smoking. Our findings are supportive of the hypotheses that glucose metabolism hyperinsulinemia, insulin resistance, and metabolic abnormalities associated with it may play a significant role in the etiology of colorectal cancer.

Hepatitis C virus (HCV) infection and liver-related mortality: a population-based cohort study in southern Italy. The Association for the Study of Liver Disease in Puglia.

BACKGROUND: Hepatitis C virus (HCV) is a common cause of chronic liver diseases but the degree to which these diseases contribute to liver-related mortality is not well established. The aim of this study was to estimate the absolute and relative effects of HCV infection on liver-related mortality. METHODS: A population random sample of 2472 subjects aged > or = 30 years was enrolled and followed up from 1985 to 1996. At enrollment, a structured interview and a clinical evaluation were performed. Serum samples were tested using HCV ELISA and RIBA HCV. Outcomes were overall and liver-related mortality and tracing procedures included review of office and hospital records, death certificates, and interviews with general practitioners, attending hospital and next of kin. Statistical analysis was performed using Poisson and binomial prospective data regression. RESULTS: Crude overall and liver-related mortality rates were 7.66 (95% CI : 6.68-8.79) and 0.9 (95% CI : 0.3-2.2) per 10(3) person-years, respectively. For HCV infection effect, incidence rate ratio and difference (per 10(3) person-year), risk ratio and difference were 27.5 (95% CI : 6.5-115.6), 4 (95% CI : 3-7), 33.1 (95% CI : 7.8- 139.3) and 0.06 (95% CI : 0.04-0.08), respectively; all measures were adjusted for age at death, sex and daily alcohol intake. CONCLUSIONS: The results show a strong relative but weak absolute effect of HCV infection on liver-related mortality in the 10-year period considered. Poisson and binomial models are virtually equivalent, but the choice of the summarizing measure of effect may have a different impact on health policy.

Cholelithiasis after Billroth II gastric resection.

We evaluate whether Billroth II (BII) gastric resection is a predisposing condition in the formation of gallstones. In our department radiologic and echographic imaging is a routine examination before abdominal surgery. From 1980 to 1985, 202 patients underwent cholecystography or ultrasonography before BII gastric resections for peptic ulcers. The prevalence of cholelithiasis in a random sample of the population of the town near our institution was used to calculate the expected frequency of gallstones. The comparison of the expected and the observed frequencies of cholelithiasis in these patients did not show any statistically significant difference between men and women. All these patients were recalled for a follow-up of the long-term effects of gastric surgery, and 66% (133/202) returned and were examined. They underwent ultrasonography of the biliary tract. Again the observed and the expected (obtained as before) frequencies of cholelithiasis were compared. A statistically significant difference was apparent in male patients. We conclude that BII gastric resection could be a predisposing factor in the formation of gallstones in men.

Influence of the method of digestive tract reconstruction on gallstone development after total gastrectomy for gastric cancer.

The aim of this study was to evaluate whether total gastrectomy performed for gastric cancer leads to an increased risk of cholelithiasis and whether the method of reconstruction of the digestive tract influences that risk. A total of 102 patients who had undergone total gastrectomy for gastric cancer between 1980 and 1990 were studied. The preoperative prevalence of cholelithiasis was 4% in men and 12% in women. Eighty-seven patients (85%) without gallstones before surgery were reexamined after gastrectomy. The postoperative prevalence of cholelithiasis in this group was 36% in men and 19% in women. Before surgery, the difference between the expected frequency of cholelithiasis (calculated on the basis of the data of a community survey) and the observed frequency was not statistically significant (p > 0.05) either in men or women. After surgery, the observed frequency of gallstones was significantly higher than the expected frequency in men (p < 0.0001) but not in women (p = 0.06). The risk of cholelithiasis was significantly higher in patients with Roux-en-Y reconstruction (n = 55) than in those with jejunal interposition (n = 32) (log-rank test, p = 0.03), and that risk was independent of age, sex, and body mass index.

Duodenogastric reflux of bile acids, gastrin and parietal cells, and gastric acid secretion before and 6 months after cholecystectomy.

In order to evaluate the effect of cholecystectomy on the gastric mucosa, the duodenogastric reflux of total and single bile acids, the number of parietal and gastrin cells, and the volume of gastric acid secretion were examined in 15 patients with gallstones and functioning gallbladders before and 6 months after cholecystectomy. The duodenogastric reflux of the total bile acids increased from a mean preoperative value of 1.9 mumol/hour to a mean postoperative value of 21 mumol/hour (p = 0.008). The duodenogastric reflux of all single bile acids increased after cholecystectomy, with a higher increase in glycoconjugated compared with tauroconjugated bile acids. The parietal cells decreased from a mean preoperative value of 82.8 to a mean postoperative value of 68.7 (p = 0.05), whereas there was only a mild increase in the number of gastrin cells; the output of gastric acid remained unchanged. The variation of the gastrin cells before and after cholecystectomy was negatively correlated only with the variation of taurocholic acid (r = -0.50, p = 0.05), while the variation of the parietal cells was mildly correlated with all single bile acids (r = 0.35-0.50, 0.05 less than p less than 0.02). These findings show an increased duodenogastric reflux of bile acids 6 months after cholecystectomy with a mild morphologic alteration of the gastric mucosa.

Epidemiology of cholelithiasis in southern Italy. Part II: Risk factors.

OBJECTIVE: To determine behavioural, dietary and other common factors associated with new cases of gallstones, diagnosed by ultrasonography, in a prospective cohort study conducted in southern Italy. SUBJECTS AND METHODS: Between May 1985 and June 1986, systematic sampling from the electoral register of Castellana, a small town in southern Italy, yielded 2472 subjects who had had their gallbladder checked for gallstones by ultrasonography. Between May 1992 and June 1993, 1962 out of the 2235 (87.7%) subjects without gallstones at baseline agreed to a further ultrasound examination. At the first survey a standardized questionnaire was administered, inquiring about medical history, diet, cigarette smoking and other behavioural characteristics. Height and weight were also measured, and blood levels of glucose, cholesterol, HDL-cholesterol and triglycerides were determined by standard methods. The same variables were measured at the second survey. The diagnosis of gallstones was made with the same echograph by echographists working in the same department. Multiple logistic regression was used to determine which factors measured at the first survey were associated with the incident cases of gallstones. RESULTS: One hundred and four subjects had developed gallstones, an incidence of 9.7 per 1000 persons per year. Age, body mass index (BMI), weight change, a history of diabetes, constipation (shown by use of laxatives), cigarette smoking, years of schooling, consumption of fried foods and excessive oil, and pregnancy in females, were positively associated with the incidence of gallstones. Consumption of wine, coffee, fish and wholemeal bread was inversely associated. Sex, family history of cholelithiasis, use of oral contraceptives and serum lipids were not independent risk factors for gallstones. CONCLUSION: The results of this study confirm many gallstone-associated factors reported in previous cross-sectional and case-control studies, as well as in other cohort studies based on the clinical diagnosis of gallstones, such as BMI, ageing and wine consumption. Furthermore, use of laxatives, considered a proxy of constipation, appears to be another important independent risk factor for gallstones.

Diet and duodenal ulcer.

BACKGROUND: Despite the fact that the main cause of duodenal ulcer incidence and recurrence is the Helicobacter pylori bacterium, more than 80% of Helicobacter pylori-infected people never develop an ulcer. Diet may be one of the most important environmental factors contributing to duodenal ulcer. AIMS: To explore the role of diet in causation, treatment and prevention of duodenal ulcer recurrence. METHODS: All research papers published in English from 1966 to October 1999 present in Medline, involving human subjects, and having duodenal ulcer as outcome, entered the review. RESULTS AND CONCLUSIONS: Soluble fibre from fruit and vegetables seem to be protective against duodenal ulcer and refined sugars a risk factor. The role of fibre in the treatment and prevention of recurrence of duodenal ulcer is uncertain, as is that of essential fatty acids. However, none of the epidemiological studies on the relationship between diet and duodenal ulcer disease controlled for Helicobacter pylori.

Cigarette smoking and ulcerative colitis. A case control study.

In order to examine the relationship between cigarette smoking and ulcerative colitis (UC), we carried out a case-control study of smoking habits in 84 patients with UC and in two different types of controls: 84 taken from a hospital population and 84 from the general population, matched to the cases for age, sex, school education and social class. Non-smokers were found to have a greater UC risk than smokers in both internal controls (Odds ratio = 4.6, 95% CL = 1.8-11.7) and external controls (OR = 10, 95% CL = 3.6-26.9). There was a larger number of ex-smokers among patients with UC than among the controls. Evaluations at the time of disease onset also demonstrated that ex-smokers had a greater relative risk. Furthermore, 84% of the ex-smoker patients had stopped smoking before the onset of symptoms. Conditional multiple logistic regression with the introduction of alcohol (wine) and coffee as additional variable factors to age, sex, education and social class confirmed the independent protective effect of smoking on UC (OR for internal controls = 4.2, 95% CL = 1.7-10.2, OR for external controls = 5.7, 95% CL = 2.3-14.1). These results indicate that non-smokers and especially ex-smokers of cigarettes have a greater risk of UC, and thus confirm the results of other studies.

Spontaneous cholelithiasis in sheep: prevalence survey and analysis of gallstones and bile.

In an abattoir survey of the consecutive series of 206 sheep at a local slaughterhouse, gallstones (concretions with a diameter equal to or greater than 1 mm) were found in the gall bladder of 24 animals. There were stones in 19 out of 146 adult sheep and in 5 out of 60 lambs, with the higher frequency in the male sex in both groups. Qualitative analysis of the stones showed that all of them were pigment (bilirubin) stones. There was no statistically significant difference (paired t test, P greater than 0.05) between the amounts of biliary cholesterol, phospholipids, bile acids and total and indirect bilirubin in 12 sheep with gallstones and 12 control sheep without gallstones, paired for age, sex and day of slaughter.

Prevalence of pigment gallstones in sheep.

In a survey of 666 sheep at a slaughterhouse, gallstones (concretions with a diameter greater than or equal to 1 mm) were found in the gallbladder of 50 sheep (7.5%), sludge (concretions with a diameter less than 1 mm) was found in 9 sheep (1.4%), and sludge plus gallstones were found in 7 sheep (1.1%). Gallstones and sludge were associated, and were more frequent in lambs and females, compared with adults and males. Qualitative analysis of the stones revealed all to be pigment (bilirubin) stones. There was a statistically significant increase of biliary bilirubin (total and indirect quota) only in sheep with gallstones plus sludge, compared with control sheep without sludge or gallstones. Concentrations of bilirubin, cholesterol, phospholipids, total and single bile aids, and total and ionized calcium were similar in the bile of sheep with gallstones, sludge, or both and control sheep. Bacteriologic analysis of the bile in 10 sheep with gallstones and 10 controls revealed bacteria in 50% of the first group and in 75% of the second group (Escherichia coli in all sheep and Salmonella spp also in 1 sheep with gallstones). These findings confirm our earlier findings of a high prevalence of black pigment gallstones in sheep. On that basis, we suggest that gallstones are associated with high total bilirubin concentration in the bile, and deconjugating bacteria are common in the biliary tract of these animals.