Reconstructing life history of the 18thcentury priest from Prozorje, Croatia: bioarchaeological and biochemical approaches

Reconstructing life history of the 18th century priest from Prozorje, Croatia: bioarchaeological and biochemical approaches | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results Reconstructing life history of the 18 th century priest from Prozorje, Croatia: bioarchaeological and biochemical approaches View ORCID Profile T. Kokotović , View ORCID Profile M. Carić , View ORCID Profile S. Stingl , View ORCID Profile M. Novak , View ORCID Profile J. Belaj doi: https://doi.org/10.1101/2025.02.04.636416 T. Kokotović a Institute of Archaeology; Jurjevska ulica 15 , 10000 Zagreb, Croatia Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for T. Kokotović For correspondence: tkokotovic{at}iarh.hr M. Carić b Centre for Applied Bioanthropology, Institute for Anthropological Research; Gajeva ulica 32 , 10 000 Zagreb, Croatia Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for M. Carić S. Stingl a Institute of Archaeology; Jurjevska ulica 15 , 10000 Zagreb, Croatia Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for S. Stingl M. Novak b Centre for Applied Bioanthropology, Institute for Anthropological Research; Gajeva ulica 32 , 10 000 Zagreb, Croatia c Department of Archaeology and Heritage, Faculty of Humanities, University of Primorska; Titov trg 5 , 6 000 Koper, Slovenia Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for M. Novak J. Belaj a Institute of Archaeology; Jurjevska ulica 15 , 10000 Zagreb, Croatia Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for J. Belaj Abstract Full Text Info/History Metrics Preview PDF Abstract The study presents a case of the adult male individual dated to the Early Modern Period buried at the church of St. Martin at Prozorje in Croatia. Based on the burial characteristic, the individual is presumed to be a member of the clergy. Using stable carbon and nitrogen isotope analysis of the dentine increments from the second permanent mandibular molar (Man 2) of the individual, this study aims to identify dietary changes during the juvenile years of the individual in order to reconstruct the timing of his admission into the seminary. Mean δ 13 C and δ 15 N values indicate mixed C3 and C4 terrestrial based diet with the visible animal protein intake. Results of the incremental dentine analysis provide more detailed information. The possible end of the weaning period is identified, followed by the diet based on C4 resources with low animal protein intake with periods of physiological stress during his early childhood. By the end of the observed period, significant dietary change is recorded characterised by the high animal protein intake and consumption of less C4 resources. According to the historical resources, this corresponds with the timing clergy cadets usually entered the seminary with the dietary regiment consisting of daily intake of meat or fish dishes accompanied by variety of fruit and vegetables (C3 resources). This is the first such study conducted in this part of Europe and it significantly contributes to our knowledge of dietary practices and certain social customs in Early Modern Period Croatia. 1. Introduction The stable isotope ratios of carbon ( δ 13 C) and nitrogen ( δ 15 N) in bone and dentine collagen have been frequently used to estimate human paleodiet. Unlike bones, which constantly remodel and represent the diet in the five to 30 years of life depending on the selected element or the age of the deceased, the primary dentine tissue does not remodel which allows us to reconstruct diet during juvenile years of the individual’s life, i.e. during the time of its formation ( Nanci, 2012 ). Recent advancements in incremental dentine analysis enabled much higher temporal resolution ( Fuller et al., 2003 ; Eerkens et al., 2011 , Beaumont et al., 2013 ). Since the timing of human dentition eruption and development is well established ( AlQahtani et al., 2010 ), microsampling of primary dentine provides a unique opportunity to track dietary changes throughout tooth’s development and recognize weaning practices ( Ganiatsou et al., 2022 ; Velte et al., 2023 ), short-term dietary changes ( Sánchez-Cañadillas et al. 2023 ), nutritional deprivation or physiological stress ( Brickley et al., 2020 ; Nicholls et al., 2020 ; Feuillâtre et al., 2022 ). This case study is the first time that incremental dentine analysis was performed on an archaeological sample from Croatia but also from a wider region, and aims to present a reconstruction of specific events in the early life of male individual form the church of St. Martin at Prozorje. Based on the archaeological context, the individual is presumed to have been a member of the clergy. By examining variations in δ 13 C and δ 15 N values of dentine sections, alongside bioarchaeological data from the skeleton and historical records, the study identifies periods of nutritional stress and dietary change, which could correspond to the individual’s admission to the seminary. 2. Materials 2.1 Archaeological site The old ruinous parish church in the town of Dugo Selo (north-western Croatia; Fig. 1 ) is located at Prozorje, at the hilltop popularly called Martin’s Hill ( Martin-Breg in Croatian), after the church’s patron saint. ‘The Land of St. Martin’ was first mentioned in a document dating back to 1209. The document was issued by Andrew II, king of Hungary and Croatia, and in it he confirms that the land in question belonged to the Knights Templar ( Dobronić, 2002 ). Since the land was named after the saint, it implies that the church was already present at this position at the beginning of 13 th century, but the oldest still preserved and visible parts of the sacral building can be dated to the second half of the 15 th century ( Belaj, 2007 ). The church of St. Martin at Prozorje was in continuous use from its erection until the very end of the 19 th century ( Košćak, 2009 ). Except for a few minor repairs during the 20 th century, a large part of the abandoned building soon collapsed. In a period from 2002 to 2018, a series of archaeological excavations were conducted at the site. Complete interior of the old church was excavated along with the two old sacristies and two lateral chapels. A narrow space along some of the outside walls, primarily those along the sanctuary, was also studied during the excavation. In total, almost 300 graves were documented, the majority of which can be dated to the 17 th and 18 th centuries ( Belaj and Stingl, 2019 ). Download figure Open in new tab Fig. 1 The map showing the location of the church of St. Martin at Prozorje (made by: T. Kokotović, 2024; source: OpenStreetMap, Creative Commons Attribution-ShareAlike 2.0 license, CC BY-SA 2.0; created with Datawrapper) 2.2 Grave no. 4 The grave no. 4 (G4) was discovered in 2002 during the first archaeological campaign. It was located in the sanctuary in front of the altar ( Figs. 2 and 3 ) and just a little bit south of the grave 2 that was covered with a secondarily used tomb slab from the period of military orders ( Belaj, 2007 ) which was placed on the most prominent position of the entire interior, at the central axis of the church. Poorly preserved remains of leather footwear were situated near both feet. The footwear was without iron fittings and buckles and most probably made with wooden fittings and tied with a lace. Along the individual’s forearms and on their chest, there were rows of still connected double looped hook-and-eye fasteners. Such fasteners first appeared in the territory of present-day Croatia at the end of the first half of the 14 th century ( Anzulović, 2007 ) and in the archaeological context, they can be found in the layers dated from the second half of the 15 th century onwards ( Predovnik et al., 2008 ). Their use rapidly spread in the 16 th and 17 th centuries ( Demo, 2007 ). Depending on the type, they were used for hooking up shirts or for coats and socks ( Azinović Bebek, 2009 ). Traces of wood and numerous finds of nails suggest that the deceased was buried in a coffin. Unfortunately, these finds cannot be used for a more precise dating of the grave. Given that this burial cut through all the preserved floors of the church and judging by the artefacts discovered in the two older burials found beneath, G4 can be dated to the 18 th century, probably to its second half ( Belaj, 2006 ; Stingl, 2024 ). Download figure Open in new tab Fig. 2 Layout of the Church of St. Martin with the marked position and orientation of G4 in the sanctuary (made by: A. Kudelić; adapted by: T. Kokotović, 2024) Download figure Open in new tab Fig. 3 G4 during the excavation (source: Archive of the Institute of Archaeology) What makes this burial different from the vast majority discovered at the site is its orientation. The orientation of the church of St. Martin at Prozorje is slightly deflected towards the SW-NE direction ( Belaj, 2007 ) and most of the burials have the same orientation as the building. To simplify, since the deflection is not that significant, we can consider the orientation of the burials as predominantly west-east, which is the most common orientation of bodies in the late medieval and post-medieval periods. When the body (with or without the coffin) was laid in the grave, the head was in the west and feet in the east. However, this individual was laid in the opposite direction, with their head in the east and the feet in the west. 3. Methods The sex and age-at-death were established using standard macroscopic methods and criteria ( Buikstra and Ubelaker, 1994 ). Possible pathological changes were diagnosed based on the criteria presented in Aufderheide and Rodríguez-Martín (1998) , and Ortner (2003) . For establishing approximate age of the formation of linear enamel hypoplasia (LEH), total crown height and CEJ-LEH 1 distance was measured for each line (Buikstra, Ubleaker, 1994) using digital calliper. Approximate age of the onset of LEH on the canines was calculated using the calculator presented by Dąbrowski et al. (2021) and the method based on datasets for medieval Northern European populations ( Reid and Dean 2006 ). This method was deemed best when considering historical and geographic context of our sample. 3.1. Faunal bulk C/N stable isotope analysis The study includes faunal material from three cows ( bos taurus ) and three pigs ( sus scrofa) from the burg Vrbovec, a medieval site adjacent to the Martin-breg. The samples are dated in the 13 th and 14 th centuries and represent the faunal baseline from carbon and nitrogen isotope ratios in the area. These are the most recent and geographically closest faunal baseline samples we were able to obtain. Collagen extraction was carried out following the modified Longin method ( Longin, 1971 ; Brown et al., 1988 ). Faunal samples were demineralized in 0.5 HCl at 4°C, then placed in a HCl solution (pH3) at 70°C for 48 hours to gelatinise. Samples were then filtered using E-zee filters and freeze dried. The collagen from faunal samples was weighed out, and the samples were then sent to the Iso-Analytical Laboratory (Crewe, Cheshire, UK). The results are reported in the conventional δ notation in the per mil (‰) relative to the international reported standards, VPDB for carbon, and AIR for nitrogen. The collagen quality of each sample was evaluated through the C/N atomic ratio and carbon and nitrogen content. 3.2. Incremental dentine analysis Stable carbon and nitrogen isotope analysis was carried out on the incremental dentine collagen from the second permanent mandibular molar (Man 2) of the studied individual. Following Method 2 from Beaumont et al. (2013) , 13 incremental dentine samples were taken from the crown to the apex of the second permanent mandibular molar (Man M2) of the individual. The tooth was first demineralized in 0.5 HCl at 4°C. Once the reaction was complete, the demineralized dentine was divided into transverse increments using a sterile scalpel, starting from the coronal dentine horn. In order for each section to produce sufficient collagen for duplicate measurements, a minimum weight after demineralization of 2.5 mg was required. The first 10 sections were divided into transverse samples at 1 mm intervals. To meet the required minimum weight of each sample, the last two increments were divided into thicker samples at 2 mm, and the final sample being divided at 5 mm. The demineralised sections were placed in a HCl solution (pH 3) at 70°C for 24 hours to gelatinise. The solution was then divided into three test tubes per sample without filtration, frozen, then freeze-dried. The collagen from human dentine samples was sent to the Isotope Ratio Mass Spectrometry Lab (SILVER) at the University of Vienna (Austria). The approximate age of each increment was based on the timing of tooth formation described in the London Atlas ( AlQahtani et al. 2010 ) for the permanent mandibular second molar. The method used in this study for calculating the approximate chronological age for each increment was adapted from Beaumont and Montgomery (2015) . The approximate age at which mandibular second molars start to develop is around 2.5 years, and the age at root completion is 15.5 years ( AlQahtani et al., 2010 ). The time it takes the tooth to form is approximately 13 years (15.5–2.5). This is then divided by the number of increments, in this case 19, considering that the last three increments in this case vary in thickness (10+2+2+5mm). Starting from the first coronal increment, this number (13/19∼0.7) is then added to the approximate age assigned for each increment. The approximate age for the last three increments was calculated by multiplying 0.7 with thickness of the increment (2 or 5mm). Statistical analysis was performed in jamovi (version 2.5) computer software for Windows (The jamovi project (2024), retrieved from https://www.jamovi.org ). The statistical test used on the data was an independent Mann-Whitney U test. 4. Results The poorly preserved remains of a complete skeleton were discovered in the G4. Bioarchaeological analysis established the individual to be a male, aged between 30 and 45 years (middle-aged adult). The teeth of this individual show evidence of caries, alveolar abscess and LEH on the maxillary and mandibular canines. Several pathological changes (slight resorption of the bone on the inferior and lateral margins of the nasal cavity, inflammatory pitting on the palatine, and porosity around the orbits) could be indicative of leprosy ( Møller-Christensen, 1961 ), but without additional changes to the postcranial skeleton and pathogen DNA analysis, at the moment this diagnosis is only speculative. The total crown height and CEJ-LEH distance for both canines, as well as the estimated approximate age of onset of LEH, are presented in Tab. 1 . Three lines were observed on each canine and the age of onset of LEH was estimated at around 2.5 years, then again around three and four years of age. View this table: View inline View popup Download powerpoint Tab. 1 Approximate age of the onset of LEH for maxillary and mandibular permanent canine of the male from grave no. 4 (made by: T. Kokotović, 2024) The results of the faunal baseline and incremental collagen analysis are shown in Tab. 2 . After the preparation of samples, all of the samples expect one (MB-4-B 10) had yielded sufficient collagen for the analysis. When human δ 13 C and δ 15 N values are compared with the faunal baseline, the expected shift in tropic level between herbivores and humans (2–5‰) is present ( Ambrose 1991 ; Hedges and Reynard 2007 ). Estimated approximate age for each increment is presented in the Tab. 3 . View this table: View inline View popup Download powerpoint Tab. 2 Isotopic data and collagen quality indicators for faunal baseline (burg Vrbovec site) and dentine sections for MB-4-B, second mandibular permanent molar (Prozorje site) (made by: T. Kokotović, 2024) View this table: View inline View popup Download powerpoint Tab 3 Approximate age of dentine sections for MB-4-B, second mandibular permanent molar (made by: T. Kokotović, 2024) The mean isotopic values for the second mandibular molar are –15.0‰ for δ 13 C and 9.3‰ for δ 15 N ( Fig. 4 ). Throughout the development of the tooth, figures display a fluctuation in both δ 13 C and δ 15 N values ( Figs. 5A , 5B and 6 ). Up to the four years of age, a slight, gradual increase is observed in δ 13 C (+0.4‰) values, followed by a decline (–1.5‰) around the age of six years. Between 10 and 12 years of age there is another gradual increase (+0.96‰) after which the δ 13 C values visibly decline (–2,4‰). δ 15 N values also fluctuate in the first 10 years of life, with an elevation in δ 15 N values (∼0.5‰) up to the age of six years, after which the values gradually increase until the age of approximately 15 years, when the recorded δ 15 N values are at their highest (11.45‰). The last two samples (MB-4-B-12 and MB-4-B-13), representing the period between the approximate ages of 11 and 15 years, significantly differ in both δ 13 C and δ 15 N values from the former period (independent sample Mann-Whitney U test p = 0.041 for both δ 13 C and δ 15 N values). Download figure Open in new tab Fig. 4 Mean δ 13 C and δ 15 N values for MB-4-B, second mandibular permanent molar (made by: T. Kokotović, 2024) Download figure Open in new tab Fig. 5A δ 13 C values of dentine sections against approximate age for MB-4-B, second mandibular permanent molar (made by: T. Kokotović, 2024) Download figure Open in new tab Fig. 5B δ 15 N values of dentine sections against approximate age for MB-4-B, second mandibular permanent molar (made by: T. Kokotović, 2024) Download figure Open in new tab Fig. 6 δ 13 C and δ 15 N values of dentine sections against approximate age for MB-4-B, second mandibular permanent molar (made by: T. Kokotović, 2024) 5. Discussion The mean δ 13 C and δ 15 N values for the individual from G4 indicate a terrestrial staple diet based on plant food with visible dairy/animal protein intake while δ 13 C values indicate a mixed diet of both C3 and C4 plants. Humans consuming a diet based on C3 plants have δ 13 C values of ∼19‰, the ones consuming a diet consisting only of C4 plants have δ 13 C values of ∼ 8‰, and individuals who consume a mixed diet of both C3 and C4 plants have δ 13 C values that are intermediate ( Dupras and Tocheri, 2007 ), as is the case here. These values correspond with the historical records from the 17 th and 18 th centuries about agricultural and husbandry activities in the region ( Adamček, 1981 ). C3 plants cultivated in Dugo Selo and surrounding regions were mainly cereals (wheat, rye, oats, barley, and pyrethrum) and vines ( Adamček, 1981 ). C4 plants included primarily millet, but also corn, whose cultivation started in the 18 th century ( Adamček, 1981 ). The animals that were raised in the region were mainly pigs, chickens, cows, and horses, with primary source of animal protein being pigs, cows, and chickens, while horses and oxen were used as draft animals ( Adamček, 1981 ). Since the second mandibular molar starts to develop around the age of 2.5 years, exclusive breastfeeding and the start of the weaning period could not be recorded. Nevertheless, the difference in δ 15 N values between the first and second samples could represent the end of the weaning process. Weaning is the period of transition from an exclusively breastmilk diet to diet combined with breastmilk, supplementary food, and drinking water, and ends with the cessation of breastfeeding ( Katzenberg et al., 1996 ). The replacement of protein from breastmilk with protein from other foods, which are generally at a lower trophic level in respect to breastmilk, results in a drop in the infant’s δ 15 N values ( Fogel et al., 1989 ). Unfortunately, there is no specific information regarding the age of cessation of breastfeeding and supplementary foods for this region in the 17 th or 18 th centuries, but ethnographic research into the rural communities in Croatia in the 19 th century reveals that the duration of breastfeeding was somewhat long, from one year of age up to the birth of the second child, meaning up to two or three years on average ( Čapo-Žmegač, 1998 ). Besides breast milk, during the weaning period, children were given easily digestible foods in the form of various porridges, made from flour, bread, or grains, especially millet (a C4 plant), diluted in milk, meat stock or water ( Pleše, 2019 ). The nutritional content of the supplementary foods varied, and subsidizing milk and stock with water would induce various metabolic disorders (scurvy, rickets) and kidney stones, and contamination of utensils and food would result in brucellosis and/or parasitic infections ( Castilho and Barros Filho, 2010 ). Therefore, the appearance of first and second LEH on mandibular canine (approximate age of the onset is at 2.5, 2.9 and 3.3 years) could be the result of the physiological stress connected to the weaning period. Several studies researching the connection between LEH and the weaning process observed the occurrence of LEH at the end or shortly after the end of complementary feeding ( Moggi-Cecchi et al., 1994 ; Sandberg et al., 2014 ; Crowder et al., 2019 ; Orellana-González et al., 2020 ). Another drop in the nitrogen and carbon values was recorded in the period between four and eight years, accompanied by the occurrence of LEH at the beginning of the observed period. The carbon and nitrogen values indicate terrestrial based diet with significant contribution of C4 resources and low animal/dairy protein intake, and in concurrence with LEH, could indicate a period of notable physiological stress. High levels of subadult stress were observed in several Early Modern Period populations from continental Croatia ( Novak et al., 2009 , 2021 ; Novak and Krznar, 2010 ; Krznar and Hajdu, 2020 ; Bedić et al., 2022 ), especially in children aged between 2 and 10 years ( Novak et al., 2009 ; Bedić et al., 2022 ). Novak and colleagues (2009) suggested that the observed physiological stress was caused by a number of various factors (parasitic infections, metabolic disorders, infectious diseases) occurring as a result of inadequate nutrition, reflecting generally poor living conditions, low health and sanitary standards in continental Croatia during this period). In our case notable changes in δ 13 C and δ 15 N values were observed after the age of 11 years: δ 13 C values are falling and δ 1 5 N values are rising. This pattern of opposing covariance, i.e. famine pattern, was recognized by Beaumont and Montgomery (2016) when studying the victims of the Great Irish Famine and interpreted it as the evidence for the period of nutritional stress prior to their death or the introduction of famine relief food. Although this explanation cannot be completely disregarded in this case, here we see gradual change over the course of approximately five years that could also indicate visible dietary change, reflecting consumption of less C4 plants and an increase in dairy/animal protein intake during the last five and half years, but especially the last two and half years of the observed period. This shift could be related to the major lifestyle change or even change in geographical location with the access to the different food resources. The location of the burial in the sanctuary of the church in front of the main altar, strongly suggests that the body was deliberately oriented east-west. The possibility that this orientation is a mistake caused by haste, as sometimes was the case ( Krznar, 2012 ), can be ruled out. A most likely explanation in this case is that this grave was the burial of a priest. After the Council of Trent (1546-1563) members of the clergy were often buried with their heads in the east facing their faithful flock ( Králíková, 2007 ). Likewise, according to the Roman Ritual of 1614, close attention is to be paid when burying members of the laity inside churches or chapels – they should have their feet towards the altar, while priests are supposed to have their head towards it ( Bradara, 2024 ). This position has its roots in the belief that, upon his second arrival, Christ will come from the east and all the dead will be facing him ( Tkalčec et al., 2021 ). For example, priest burials with this orientation were excavated in the Minster Church in Bonn ( Keller, 2000 ) and one was documented in the filial church of St. Bartholomew in Martkt Indersdorf ( Mittelstraß, 2007 ), both in Germany. All the graves with an east-west orientation near the main altar in the church of St. Wenceslas in Ostrava, Czechia are assumed to belong to the clergy ( Králíková, 2007 ). Two graves with this orientation were excavated in front of the altar in the parish church of the Assumption of Mary in Hollenburg, Austria. Both are interpreted as priest burials, and for one of those two we can say this without a doubt since most parts of the vestment of the deceased could be identified: a cassock, an alb, a cingulum, a chasuble, a stole, and a maniple, revealing that he was, indeed, a priest ( Leib, 2008 ; Grömer, 2016 ). If we presume that the individual from G4 in Prozorje was indeed a member of the clergy, could a significant change in diet reflected in changes in his N/C isotopic values reflect the time when he entered the seminary? During the Council of Trent, the Decree on the establishment of the episcopal seminaries was passed. The Decree highlights a set of rules and regulations regarding the upbringing and education of clergy, prescribing the minimum age of 12 for entering the seminary ( Škreblin, 1944 ) which corresponds with the timing of the dietary changes observed in our sample. After the Council, episcopal seminaries were established across Christian Europe in accordance with the regulations laid out by the Decree ( Škalabrin, 2006 ). Seminary in Zagreb was established in the second half of the 16 th century ( Patafta, 2020 ), and in Varaždin in 1659 ( Puhmajer, 2011 ). Based on the available historic sources regarding the diet of the clergy cadets, it seems that their meals included substantial quantities of meat and fish that were eaten on the regular basis. According to the regulations of the Flori’s seminary in Zadar, staff and the cadets dined together and shared the same meal ( Dundović, 2020 : 104). They were given three meals a day (breakfast, lunch and dinner): for breakfast, they served a piece of bread with a glass of vine every day, and some fruits and cheese once a week while lunch usually consisted of soup and one meat or fish meal, depending on the day, with the addition of fruits and cheese after the main course ( Dundović, 2020 ). Dinner included salad or soup, after which, the fruit and cheese were again consumed ( Dundović, 2020 ). The prescribed amount of meat and/or fish amounted to 12 ounce per day/per person. Meat consumed in this seminary included beef and lamb, but not so much veal and poultry because of its scarcity in Zadar and the surrounding region ( Dundović, 2020 ). It has to be noted that the cadets from Flori’s seminary were members of the nobility and urban population living on the Adriatic coast, so this description cannot be literally transcribed into the geohistorical context of continental Croatia. However, in the seminary in Đakovo in eastern Croatia, established at the beginning of the 19 th century, cadets were given two meals: lunch at 11 AM and dinner at 6:30 PM ( Sršan, 1996 ). The cook had to prepare four different meals for lunch and three for dinner ( Sršan, 1996 ). The meals often contained roasted meat with a side dish, and beef had to be prepared a day earlier; food had to be well cooked and clean ( Sršan, 1996 ). Most bulk carbon and nitrogen isotopic studies examining the diets of monastic communities in medieval and early modern Europe have primarily focused on the dietary patterns of adult members of the clergy. Studies on incremental dentine analysis examining the dietary patterns of monastic populations are limited, making this research a valuable contribution to the existing body of knowledge on the subject. The diet of these monastic communities generally resembled that of the contemporary nobility more than that of the common population. It was predominantly based on C3 plant resources and included a high intake of animal protein, as well as freshwater and marine resources ( Polet and Katzenberg, 2003 ; Müldner and Richards, 2005 ; Müldner et al., 2009 ; Yoder, 2012 ; Quintelier et al., 2014 ; Simčenka et al., 2020 ; López-Costas et al., 2021 ; Carić and Novak, 2023 ). Several incremental dentine studies have investigated the dietary changes of medieval and postmodern monastic populations in Europe. Kancle et al. (2018) investigated dietary changes within a friar population from two medieval friaries in northern England aiming to identify shifts in diet upon individuals’ entry into the religious order. Their analysis revealed a transition from a diet primarily based on terrestrial protein during early childhood to one that incorporated more marine protein by early adulthood. While the age at which these dietary changes occurred varied among individuals, the authors concluded that these changes corresponded to the period when the individuals entered the religious order. Similar study was conducted by Pliego and Beaumont (2023) at the monastery of San Milàn de la Cogolla Yuso, Spain during the 17 th and 18 th century. Their research identified distinct dietary patterns during the formative years of the monks, prior to their entry into the religious order, with some consuming lower-status foods and others higher-status foods. Following their induction, however, the monks adopted a more homogenous dietary regimen characterized by the consumption of C4 resources and a high intake of meat ( Pliego and Beaumont, 2023 ). 6. Conclusion The incremental dentine δ 13 C and δ 15 N profile of the male individual from the church of St. Martin at Prozorje, Croatia reveals significant insights into the dietary patterns during juvenile years of the individual in question. The isotopic values indicate a primarily terrestrial diet, including both C3 and C4 plants, alongside a noticeable intake of animal protein at the end of the profile. The shifts in isotopic values correlate with key stages during childhood, such as the end of the weaning process, as well as periods of physiological stress, potentially caused by poor nutrition, parasitic infections, or metabolic disorders, which were common in early modern period Croatia. The isotopic evidence also suggests significant dietary change after the age of 10, characterized by a reduction in C4 plant consumption and an increase in animal protein intake, which may reflect his entry into the seminary. This is supported by the burial orientation and historical context, which suggest that the studied individual was most probably a member of the clergy, and clergy cadets were known to consume diets rich in meat and fish. The findings of this study are in accord with similar studies on monastic diets in medieval and post-medieval Europe, reinforcing the idea that entering religious life often involved a transition to a diet resembling that of the higher social classes, with a greater consumption of animal protein. Overall, this study contributes to our understanding of dietary practices in Early Modern Period Croatia and the complex relationship between food, health, and social status in this period. Funding This work has been supported by the Croatian Science Foundation under the project Development and Heritage of the Military Orders in Croatia (milOrd) (HRZZ, IP-2019-04-5513). Footnotes ↵ 1 CEJ-LEH – distance between cementoenamel junction (CEJ) and the LEH References cited ↵ Adamček , J ., 1981 . Povijest vlastelinstva Božjakovina i okolice , Kaj. XIII(IV) , 105 – 134 . ↵ AlQahtani , S.J. , Hector , M.P. , Liverside , H.M ., 2010 . 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