Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: This research study consists of three investigations with regard to ostrich (Struthio camelus var. domesticus) production, meat quality and the processing of ostrich meat into a value added meat product. The first study was conducted in order to establish whether the gradual replacement of soybean oilcake meal with cottonseed oilcake meal (CSOCM) as a protein source in the diet of slaughter ostriches would affect ostrich growth performance and meat quality. A total of 105 ostriches were divided into five feeding groups according to the CSOCM inclusion level: Control (0% CSOCM), 3%, 6%, 9% and 12% CSOCM, and fed with experimental diets from 6 to 13 months of age. As a result of feeding CSOCM, the final live weight and the average daily gain significantly increased in the 12% CSOCM group compared to the other treatment diets. The proximate composition, cholesterol content, mineral and fatty acid profile of the meat remained unaffected. Considering all the results, CSOCM may be used as an alternative protein source to soybean oilcake meal in ostrich nutrition, resulting in decreased feed costs. Secondly, a descriptive sensory analysis, together with chemical and physical measurements, was performed to determine whether the manipulation of the fatty acid composition in the fan fillet (Iliofibularis muscle) as a result of feeding CSOCM would be detected on a sensory level. Two levels of CSOCM were investigated; 0% as a control and 9% CSOCM. No significant differences were found for the physical measurements (cooking loss (%) and shear force) as well as for the pH and proximate composition of the raw fan fillet. The Control group presented a higher (P<0.05) mono-unsaturated fatty acid (MUFA) content in the cooked fan fillet whereas the 9% CSOCM group showed a favourable increased (P<0.05) poly-unsaturated fatty acid (PUFA) content when compared to the cooked Control samples. As a result, the poly-unsaturated:saturated fatty acid (PUFA:SFA) ratio in the 9% CSOCM group was also higher (P<0.05). No differences (P>0.05) were found between the treatments for the n-6:n-3 (omega 6 to omega 3) ratio. The 9% CSOCM group had a more intense beef aroma, had a higher level of initial and sustained juiciness as well as increased tenderness (P<0.05). Inclusion of 9% CSOCM resulted in a favourable cooked ostrich fan fillet. Finally, the effect of feeding CSOCM on a processed ostrich meat product was investigated. Fan fillet (Iliofibularis muscle) from 13 month old birds receiving no cottonseed oilcake meal (Control) or 9% cottonseed oilcake meal (9% CSOCM) was used. Olive oil was used as a replacement for pork fat, and warthog (Phacochoerus africanus) meat was used to replace commercial pork meat in the production of a semi dry sausage, cabanossi. Olive oil was included at three levels (0%, 1% and 2%). Six treatments were investigated: Control 0% olive oil, Control 1% olive oil, Control 2% olive oil, 9% CSOCM 1% olive oil, 9% CSOCM 2% olive oil en 9% CSOCM 2% olive oil. The Control and 9% CSOCM ostrich meat did not differ significantly in chemical composition nor fatty acid profile. After smoking and drying the fat content in the cabanossi containing 0%, 1% and 2% olive oil averaged 7.2%, 7.45% and 8.65% respectively. Processed meat products containing less than 10% fat are classified as a low-fat meat product. Olive oil is a mono-unsaturated vegetable oil containing mainly Oleic acid (C18:1n9c), and low quantities of saturated fatty acids and polyunsaturated fatty acids. Total mono unsaturated fatty acids in the cabanossi increased from 47.0% to 73.0% of total fat, whilst total saturated fatty acids and total polyunsaturated fatty acids decreased from 40.6% to 19.9% and 11.6% to 6.6% respectively as olive oil increased from 0% to 2%. The inclusion of olive oil at 2% resulted in cabanossi with increased (P<0.05) tenderness, juiciness and cured red meat colour, all factors that appeal greatly to the consumer. Overall flavour was not adversely affected by the inclusion of olive oil. This investigation indicated that the use of CSOCM had no negative effect on the production performance of ostriches whilst a 9% CSOCM inclusion level resulted in meat that was found to be favourable by a trained sensory panel. Furthermore, the use of CSOCM as a feed component also had no negative effect on a processed product (cabanossi) derived from the meat obtained from the birds fed this feed component. The CSOCM used in this investigation had low levels of gossypol (10 to 20ppm) and more research is required on the effect of the use of CSOCM with higher levels of gossypol on the production performance of ostriches. / AFRIKAANSE OPSOMMING: Die studie het bestaan uit drie ondersoeke met betrekking tot volstruis (Struthio camelus var. domesticus) -produksie, -vleiskwaliteit en die vervaardiging van waarde-toegevoegde geprosesseerde volstruis-vleisprodukte. Die doel van die eerste studie was om vas te stel of die geleidelike vervanging van sojaboonoliekoekmeel met katoensaad-oliekoekmeel (CSOCM) as ‘n proteïenbron in die voeding van volstruise, die groeipersentasie en vleiskwaliteit van die Iliofibiularis spier (fan fillet) sal affekteer. ‘n Totaal van 105 volstruise is verdeel in vyf voedingsgroepe volgens die katoensaad oliekoekmeel insluitingsvlak: Kontrole (0% CSOCM), 3%, 6%, 9% en 12% CSOCM. Die onderskeie voedingsgroepe was van ses tot 13 maande ouderdom op die eksperimentele voere geplaas. Die resultate het aangedui dat die voëls in die 12% CSOCM behandelingsgroep ‘n betekenisvolle (P<0.05) toename in finale lewende massa asook gemiddelde daaglikse toename gehad het. Die proksimale samestelling, cholesterol-inhoud, mineraal- en vetsuursamestelling van die vleis was nie geaffekteer deur die insluiting van CSOCM nie. Die CSOCM kan dus wel as ‘n alternatiewe proteïenbron in die voeding van volstruise gebruik word. Laasgenoemde bevinding kan ook lei tot verlaagde voerkostes, aangesien CSOCM heelwat goedkoper is as sojaboon-oliekoekmeel. Die tweede deel van die studie was van ‘n chemiese asook sensoriese aard. ‘n Beskrywende sensoriese analiese is uitgevoer om vas te stel of die manipulering van die vetsuursamestelling in die volstruis fan fillet as gevolg van die CSOCM sensories waargeneem kan word. Die chemiese en fisiese eienskappe van die vleis is ook ondersoek. Twee vlakke van CSOCM inhoud is ondersoek; 0% (as kontrole) en 9% CSOCM. Geen betekenisvolle verskille is gevind vir die fisiese vleiskwaliteit (kookverliespersentasie en taaiheid), asook vir die proksimale samestelling en pH van die fan fillet nie. Die gekookte fan fillet van die Kontrole behandeling het ‘n betekenisvolle (P<0.05) toename in mono-onversadigde vetsure (MUFA) getoon en die 9% CSOCM het ‘n voordelige toename in poli-onversadigde vetsuur-inhoud (PUFA) gehad. Die poli-onversadigde tot versadigde vetsuurverhouding (PUFA:SFA) was as ‘n gevolg ook betekenisvol hoër. Geen verskille (P>0.05) is opgemerk in die omega-6 tot omega-3 poli-onversadigde vetsuurverhouding (n-6:n-3) nie. Met betrekking tot die sensoriese eienskappe het die 9% CSOCM ‘n meer opvallende beesvleis aroma, hoër vlakke van aanvanklike sappigheid en ook sagter vleis in vergelyking met die kontrole behandeling gehad (P<0.05). Insluiting van 9% CSOCM het gelei tot ‘n gekookte volstruis fan fillet van voornemende kwaliteit. Laastens is daar ondersoek ingestel op die vervanging van varkvet met olyfolie in die vervaardiging van ‘n volstruis cabanossi. Chemiese asook sensoriese analises is uitgevoer op die gedroogde en gerookte volstruis cabanossi. Vir die vervaardiging van laasgenoemde produkte is die fan fillet van 13 maande oue voëls van die Kontrole (0% CSOCM) en 9% CSOCM behandelings gebruik. Addisioneel tot die volstruisvleis is daar ook vlakvarkvleis (Phacochoerus africanus) gebruik om die kommersiële varkvleis te vervang. Olyfolie was ingesluit teen drie vlakke (0%, 1% en 2% van die totale mengsel). Ses behandelings was ondersoek: Kontrole 0% olyfolie, Kontrole 1% olyfolie, Kontrole 2% olyfolie, 9% CSOCM 1% olyfolie, 9% CSOCM 2% olyfolie en 9% CSOCM 2% olyfolie. Daar was geen verskille (P>0.05) in die chemiese en vetsuursamestellings van die Kontrole en 9% CSOCM volstruisvleis nie. Na die droging en rooksiklus was die gemiddelde vet-inhoud van die 0%, 1% en 2% olyfolie cabanossi monsters onderskeidelik 7.2%, 7.45% en 8.65%. Geprosesseerde vleisprodukte met ‘n vet-inhoud van minder as 10% word in die kommersiële vleisindustrie na lae vet vleisprodukte verwys. Olyfolie is baie ryk aan MUFA, veral Oleïensuur (C18:1n9c) en dit bevat ook lae hoeveelhede SFA en PUFA. Die totale MUFA inhoud in die cabanossi het toegeneem van 47.0% tot 73.0% terwyl die totale SFA en PUFA onderskeidelik afgeneem het van 40.6% tot 19.9% en 11.6% tot 6.6%, met ‘n olyfolie toename van 0% tot 2%. Die insluiting van olyfolie teen 2% het gelei tot ‘n sagter cabanossi wat meer sappig was met ‘n meer opvallende rooi gekuurde vleiskleur, wat almal eienskappe is wat dié produk meer aantreklik maak vir die verbruiker. Hierdie studie het aangedui dat CSOCM geen negatiewe effek gehad het op die produksie van volstruise nie. Volstruisvleis van die behandelingsgroep wat CSOCM teen 9% van die dieet ontvang het, het wel vleis geproduseer wat as aanvaarbaar aanskou was deur ‘n opgeleide sensoriese paneel. Die gebruik van CSOCM as ‘n voerbestandeel het ook geen negatiewe effek gehad op ‘n geprosesseerde produk (cabanossi) gemaak van die volstruisvleis nie. Die CSOCM wat in die huidige studie gebruik is, het baie lae vlakke van gossypol (10 – 20dpm) gehad en verdere ondersoek is noodsaaklik om die effek van CSOCM met hoër vlakke van gossypol op die produksie van volstruise te bevestig.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The ostrich industry is a predominantly quantitative industry; focused mainly on the production of large numbers of slaughter birds for maximum meat and leather yield. Competing in the international market in the current economic environment necessitates a more qualitative approach. Productivity and product quality are aspects that need to be improved in order to stay competitive and economically viable. Genetic parameters for ostrich slaughter traits are lacking, however, and breeding programs are yet to be developed. Data on quantitative and qualitative production and slaughter traits from a commercial ostrich breeding flock was consequently analysed to establish the relative importance of genetic and non-genetic influences on these traits. Genetic and environmental (co)variances as well as estimates of heritability, genetic and phenotypic correlations were estimated for and among the various traits using standard software for multi-trait genetic analyses. Substantial variation, high and favourable genetic correlations as well as moderate to high heritability estimates were found among, and for distinguished body weight traits of growing ostriches. Heritability estimates of 0.14, 0.22, 0.33, 0.43 and 0.43 for 1-month, 4-month, 7-month, 10-month and 13-month-old ostrich weights were estimated in a five-trait animal model analysis. All carcass component weight traits, with the exception of the weight of the liver, showed significant genetic variation. No significant maternal permanent environmental variance was evident for these traits. Heritability estimates ranged from 0.21 (for subcutaneous fat weight) to 0.45 (for neck weight) in multi-trait analyses. The only potentially unfavourable correlation was a high genetic correlation between live weight and subcutaneous fat weight, as fat is considered as a waste product in the present system. The heritability estimates for individual muscle weights ranged from 0.14 to 0.43, while the genetic correlation between these weights and pre-slaughter live weight were all positive, ranging from 0.59 to 0.82. When meat quality traits were analysed it was evident that lightness (L*) and ultimate pH (pHu) showed significant genetic variation, with heritability estimates of 0.37 and 0.42, respectively. L* and pHu were negatively correlated (-0.65 ± 0.19). Since pH is an indicator of various meat quality parameters, it could be considered as an appropriate selection criterion for enhanced meat quality. With the exception of skin grading and crown length, all quantitative and qualitative skin traits showed significant genetic variation. Nodule traits were accordingly moderate to highly heritable. A negative, but favourable, correlation between weight and hair follicle score was ascertained, as hair follicles is a defect that should be selected against. This study demonstrated that sufficient genetic variation exists for most slaughter traits to allow sustained genetic progress for these traits, should it be desired as part of the overall selection objective. Combining some of the current economically important slaughter traits in a provisional selection index, it was clear that weight and crust skin size contributed most to monetary gain (approximately 54 and 38%, respectively). It was also demonstrated with this simple index that monetary gains in slaughter bird production should be easy to achieve at all levels of production performance and data recording. / AFRIKAANSE OPSOMMING: Die volstruisbedryf is hoofsaaklik ‘n kwantitatiewe bedryf wat meerendeels fokus op die produksie van groot getalle slagvolstruise vir die produksie van vleis en leer. Siende dat die bedryf hoofsaaklik op uitvoere fokus, word aanvaar dat ‘n verandering in strategie na ‘n meer kwalitatiewe benadering nodig is, in ag geneem die huidige ekonomiese situasie en marktoestande. Produktiwiteit sowel as produkgehalte moet in ag geneem word vir die bedryf om lewensvatbaar te bly. Daar is egter ‘n gebrek aan genetiese parameters vir volstruisslageienskappe, terwyl doeltreffende teeltstelsels nog ontwikkel moet word. Data van kwantitatiewe en kwalitatiewe produksie- en slageienskappe is gevolglik van ‘n kommersiële volstruis teeltkudde verkry en ontleed om die relatiewe belang van genetiese en nie-genetiese effekte op die eienskappe te kwantifiseer. Genetiese- en omgewings (ko)variansies, asook beramings van oorerflikheid sowel as genetiese en fenotipiese korrelasies, is vervolgens vir en tussen die onderskeie eienskappe beraam deur van standaard sagteware vir veelvuldige-eienskap genetiese ontledings gebruik te maak. Aansienlike variasie, hoë en meestal gunstige korrelasies, sowel as matige tot hoë oorerflikhede, is tussen en vir die onderskeie ligaamsgewigte van groeiende volstruise gevind. Oorerflikheidsberamings van 0.14, 0.22, 0.33, 0.43 en 0.43 is vir 1-maand, 4-maande, 7-maande, 10-maande en 13-maande-oue volstruise in ‘n vyf-eienskap dieremodel ontleding gekry. Alle karkaskomponentgewigte, met die uitsondering van die gewig van die lewer, het betekenisvolle genetiese variasie getoon. Oorerflikheidsberamings het tussen 0.21 (vir onderhuidse vetgewig) en 0.45 (vir nekgewig) gevarieer in veelvuldige-eienskapontledings. Die enigste moontlike ongunstige korrelasie was tussen liggaamsgewig en onderhuidse vetgewig, siende dat vet as ‘n afvalproduk gereken word in die huidige stelsel. Die oorerflikhede van die gewigte van indiwiduele spiere het van 0.14 tot 0.43 gevarieer, terwyl die genetiese korrelsies tussen hierdie gewigte en voorslaggewig deurgaans positief was, met waardes wat van 0.59 tot 0.82 gewissel het. Tydens die ontleding van vleisgehalte eienskappe was dit duidelik dat ligtheid (L*) en uiteindelike pH (pHu) genetiese variasie getoon het, met oorerflikheidsberamings van onderskeidelik 0.37 en 0.42. L* en pHu was negatief gekorreleerd op die genetiese vlak (-0.65 ± 0.19). Aangesien pH ‘n aanduiding is van verskeie vleisgehalteparameters, kan dit moontlik as ‘n indirekte seleksie-kriterium vir verbeterde vleisgehalte gesien word. Alle kwantitatiewe en kwalitatiewe veleienskappe het genetiese variasie getoon, met die uitsondering van velgradering en kroonlengte. Knoppie-eienskappe van die veerfollikels op die vel was ooreenstemmend matig tot hoog oorerflik. ‘n Negatiewe, maar gunstige, genetiese korrelasie is tussen liggaamsgewig en haarfollikelpunt beraam, siende dat haarfollikels ‘n defek is waarteen daar geselekteer moet word. Hierdie studie dui op voldoende genetiese variasie vir die meeste slageienskappe om voldoende genetiese vordering te verseker indien dit verlang sou word. Somminge van hierdie eienskappe wat tans van ekonomiese belang is, is vervolgens in ‘n voorlopige seleksie-indeks gekombineer. Dit was duidelik dat liggaamsgewig en velgrootte die meeste tot monetêre vordering bygedra het (onderskeidelik ongeveer 54 en 38%). Dit is vervolgens aangetoon dat monetêre vordering maklik haalbaar behoort te wees op alle vlakke van produksieprestasie en data-aantekening.

Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Although transport and lairage of ostriches are accepted causes of production losses, these losses have not yet been quantified. Transport and lairage regulations focus on the wellbeing of the birds and, by default reduce some losses. This thesis investigated weight losses and meat quality of ostriches as a result of transport density, lairage duration and lairage feed availability (ad libitum). All birds were reared on the same farm and loaded and transported together. They were randomly divided into their respective groups on loading for the transport trial, and on offloading for the lairage trial. Bird grouping was according to density for the transport trial (4 groups, H1, H2: 0.56m2/bird and L1, L2: 0.96m2/bird) and according to time spent in lairage and feed availability for the lairage trial (n=30 birds/group; L0hr and L24hr; n=15 birds/group L48hr and L48hr ad libitum feed). Behavioural observations of the ostriches showed a tendency of the ostriches to lean against objects and to orientate towards forces exerted on it to help keep its balance. Reactions to sound fluctuations were noted, with birds reacting towards changes in sound volume during transport and lairage. Timepoint numbers were allocated for each time the birds were weighed during the trial. Time points 1, 2, 3, 4 and 5 were allocated to loading, arrival, 19 hr in lairage, 31 hr in lairage and 39 hr in lairage respectively for Trial 2. Results showed no differences (P > 0.05) in live weights or meat quality parameters between Groups H1, H2, L1 and L2 during transport. Differences (P < 0.05) were found in cumulative weight losses between L0hr and the rest of the groups for time point 1. Differences between L48hr and L48hr ad libitum were found for time point 4 for cumulative weight loss. L48hr also differed significantly between the other lairage duration groups for dressing percentages as a function of loading weight. Ad libitum feed availability had a significant effect on live body weight changes but not the meat quality parameters for the groups held for 48hr in lairage. The number of birds having bruises (≈50% per group) was similar between groups and lairage had no influence on bruising. Results seem to indicate that the evaluated transport densities had no effect on the weight loss or meat quality of ostriches. However, the results indicate that the lairage period should be studied further with specific reference to weight losses during lairage. Meat quality was unaffected by the lairage parameters reported in this thesis. / AFRIKAASE OPSOMMING: Hoewel vervoer en voorslag hou (“lairage”) aanvaar word as die oorsake van produksieverliese, is hierdie aspekte nog nie gekwantifiseer nie. Regulasies in terme van vervoer en voorslag hou fokus op die welstand van die voëls en gevolglik verminder sommige verliese met hierdie faktore. Konflik tussen boere en sekondêre produsente (abattoirs en verwerkers) oor die kostes verbonde aan hierdie gewigsverliese, is in wese ‘n oorsaak van oorbruggingsfases (vervoer). Daarom is die voorslag hou-parameters van tyd, voerbeskikbaarheid en laai digtheid ondersoek. Twee proewe is geloods; een vir vervoer en een vir voorslag aanhou. Voëls is voor hulle op vragmotors gelaai is, geweeg en op spesifieke tye daarna geweeg. Diere van die laaidigtheidsproef is ingedeel in vier groepe (H1, H2: 0.56m2/voel en L1, L2: 0.96m2/voel). Met aflaai is die eerste toets groep geslag terwyl die tweede toets groep ingedeel is volgens die tyd gespandeer en voer beskikbaarheid in voorslag aanhou (n=30 voëls/groep - Groepe L0u, L24u; n=15 voëls/groep - L48u en L48u ad libitum gevoer). Gedrag van voëls is aangeteken tydens vervoer. Finale gewig was slagpale gewig en elke tyd periode waar die voëls geweeg is, is aangeteken as ‘n tydpunt. Vleiskwaliteit analises is gedoen op die linker boud se fan filet. ‘n Standaard rak leeftyd toets is ook gedoen om kwaliteit parameters te toets oor tyd. pH is gekatorigiseer volgens groepe om moontlike verskille te beklemtoon. Gedrag is opgeteken volgens die oriëntasie en steunings gewoontes van die volstruise gedurende vervoer. Voëls probeer hulle balans te hou deur op voorwerpe en op mekaar te leun, asook om hul liggaam oriëntasie te gebruik. Reaksies tot klank veranderinge is ook opgemerk tydens vervoer en voorslag aanhou. Resultate toon geen betekenisvolle verkille tussen digtheid groepe H1, H2, L1 en L2 nie. Geen gewigsverskille is gevind tussen vervoerdigtheid groepe nie. Vleiskwaliteit-parameters is ook ondersoek en geen verskille is gevind tussen groepe nie. Resultate toon wel betekenisvolle verskille in gewigsverlies-persentasie tussen groepe vir tyd in voorslag aanhou. Groepe L48u en L48u ad libitum, het onder andere betekenisvol verskille getoon vir tydpunt 4. Betekenisvolle verskille in uitslagpersentasie as funksie van begin-gewig tussen 48 uur en die res van die groepe is ook gevind. Verdere vleiskwaliteit-parameters (drupverlies, pH, ens.) tussen groepe het geen betekenisvolle verskille getoon nie. Resultate dui aan dat voer beskikbaarheid het ‘n invloed op gewigsverskille in voorslag aanhou. Regulasies vir voorslag hou, vervoer en welstand van volstruise sal moontlik verder ondersoek moet word met spesiale klem op die nut van voorslag aanhou en die se invloed op gewigsverlies. Vleiskwaliteits-parameters is nie beïnvloed deur die aspekte ondersoek nie.

Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Mycoplasma nasistruthionis sp. nov. str. Ms03 (Ms03) is one of three Mycoplasma species that were identified from ostriches. Mycoplasmas infections have been implicated in ostrich chick mortalities, growth retardation and downgrading of ostrich carcasses. Currently there is no vaccine available for the treatment of mycoplasmosis in ostriches. This study investigated the development of DNA vaccines against Ms03 infections in ostriches. To this end, the Ms03 genome was sequenced and annotated. The vaccine candidate gene, oppA, was identified within the genome sequence and characterized before DNA vaccines containing the oppA were developed and tested. The genome of Ms03 was sequenced and the resulting 172 contigs were annotated. This dissertation presents the first Ms03 draft genome and annotation which contributed to the understanding of Ms03 as a miniature genetically independent organism. In Ms03, genome replication, cell division, RNA transcription, protein translation and glycolysis resemble that of the closely related Mycoplasma synoviae 53. Purine and pyrimidine metabolism was incomplete and de novo synthesis thereof was not possible. Amino acid synthesis in Ms03 was mostly absent and only the genes that convert aspartate to asparagine and glycine to serine were found. More importers than exporters were annotated owing to the lack of synthesis pathways in Ms03, which is typical for mycoplasmas that have parasitic life styles. Two oligopeptide permease (opp) operons were annotated within the Ms03 genome. The potential of the oppA as a vaccine candidate gene was evaluated by investigating the need for a substrate-binding domain (OppA) as part of the OppBCDF transporter within Mycoplasma species. An oppA homologue could be identified for each oppBCDF operon in all species and therefore must play an essential role in oligopeptide transport. All mycoplasmas (except for hemoplasma) had one, two or three opp operons that could be divided into three types (Type A, B and C). Each type had unique InterPro and MEME domains and motifs which together with the phylogenetic analysis suggest unique roles in their survival under different conditions. Ms03 had a Type A and a Type B opp operon, the Type A oppA was used as vaccine candidate gene. The Type A oppA was cloned and site-directed mutagenesis was used for codon correction before the mutated gene was sub-cloned into three DNA vaccine vectors. The three DNA vaccines (pCI-neo_oppA, VR1012_oppA and VR1020_oppA) were used to vaccinate ostriches and the OppA-antibody response was analysed by ELISA. The VR1020_oppA and pCI-neo_oppA constructs elicited a primary immune response in ostriches, indicating that the OppA protein was expressed in vivo and was immunogenic. This can therefore be viewed as the first step in the development of a DNA vaccine for the control of mycoplasma infections in ostriches. / AFRIKAANSE OPSOMMING: Mycoplasma nasistruthionis sp. nov. str. Ms03 (Ms03) is een van drie mikoplasma spesies wat volstruise infekteer. Mikoplasma-infeksies in volstruise veroorsaak kuiken vrektes, vertraagde groei en afgradering van volstruis karkasse. Daar is tans geen geregistreerde mikoplasma entstof beskikbaar vir gebruik in volstruise nie. Hierdie studie het die ontwikkeling van DNS-entstowwe teen Ms03-infeksies in volstruise ondersoek. Vir hierdie doel was die Ms03-genoomvolgorde bepaal en geannoteer. Die entstofkandidaat-geen, oppA, was geïdentifiseer in die genoomvolgorde en gekarakteriseer voordat DNS-entstowwe (wat die oppA-geen bevat) ontwikkel en getoets is. Die Ms03-genoomvolgorde was bepaal en die gegenereerde 172 aaneenlopende volgordes was geannoteer. Hierdie proefskrif bied die eerste voorlopige volgorde en annotering van die Ms03-genoom wat bygedra het tot die kennis van Ms03 as 'n miniatuur geneties onafhanklike organisme. Genoom-replikasie, seldeling, RNS-transkripsie, proteïen-translasie en glikolise in Ms03 stem ooreen met dié prosesse in die naverwante Mycoplasma synoviae 53. Die purien en pirimidien metabolisme was onvolledig en de novo sintese daarvan was nie moontlik in Ms03 nie. Aminosuursintese in Ms03 was meestal afwesig en net die gene wat aspartaat omskep na asparagien en glisien na serien was gevind in die annoteerde genoom. Meer invoerders as uitvoerders was geannoteer, wat dui op die gebrek aan sintesepadweë in Ms03. Dit is tipies van mikoplasmas wat ‘n parasitiese lewensstyle het. Twee oligopeptied-permeases (opp) operons was gevind in die Ms03-genoom. Die potensiaal van die oppA-geen as 'n entstofkandidaat-geen was geëvalueer deur die behoefte van 'n substraatbindingsdomein (OppA) as deel van die OppBCDF-vervoerder binne mikoplasma spesies te ondersoek. 'n Homoloog van die oppA-geen kon geïdentifiseer word vir elke oppBCDF-operon in al die spesies en behoort daarom 'n noodsaaklike rol te speel in die vervoer van oligopeptiede. Alle mikoplasmas (behalwe vir hemoplasmas) het een, twee of drie opp-operons, wat verdeel kan word in drie tipes (Tipe A, B en C). Elke tipe het unieke InterPro en MEME domeine en motiewe wat saam met die filogenetiese ontleding daarop dui dat hulle unieke rolle in oorlewing onder verskillende omstandighede speel. Ms03 het 'n Tipe A en Tipe B opp-operon, die Tipe A oppA is gebruik as entstofkandidaat-geen. Die Tipe A oppA-geen was gekloneer en teikengerigte-mutagenese was gebruik vir kodonregstellings voordat die gemuteerde geen in drie DNS-entstof vektore gesubkloneer was. Die drie DNS-entstowwe (pCI-neo_oppA, VR1012_oppA en VR1020_oppA) was gebruik om volstruise in te ent en die OppA-teenliggaamsreaksie was geanaliseer deur ELISA. Inenting met die VR1020_oppA en pCI-neo_oppA entstowwe het tot 'n primêre immuniteitsreaksie in volstruise gelei. Dit dui daarop dat die OppA proteïen in vivo uitgedruk en immunogenies was. Dit kan beskou word as die eerste stap in die ontwikkeling van 'n DNS-entstof vir die beheer van mikoplasma-infeksies in volstruise.

Hoje a criação de avestruz ((Struthio camelus, Linnaeus 1758)) é uma atividade de grande potencial, porém não existem padrões definidos sobre a histologia do seu fígado, que é um órgão de grande importância no metabolismo, o conhecimento de sua histologia e anatomia pode contribuir para a detecção de doenças e deficiências nutricionais que influenciam no crescimento e desenvolvimento do animal. Os objetivos desta pesquisa são: Estudar a anatomia e histologia do fígado e a ramificação de sua artéria hepática, veia porta hepática e ducto biliar. Para a realização da parte macroscópica foram utilizados quinze avestruzes com idades entre 12 e 18 meses (com peso médio em torno de 80 a 100 kg), provenientes do abatedouro Don Pig, situado próximo à cidade de Botucatu no estado de São Paulo. Os animais foram abatidos com pistola pneumática e posteriormente submetidos a sangria. Foram injetadas com látex a artéria hepática, o ducto biliar e a veia porta. O fígado dos avestruzes apresentam dois lobos (direito e esquerdo). No caso o direito é maior que o esquerdo e ambos são subdivididos em dorsal, intermédio e ventral. Além disso, amostras do fígado foram processadas para a observação em microscopia de luz e microscopia eletrônica de transmissão (MET). A hematoxilina e eosina (H.E), picrossírius, Gordon e Sweets, Sudan black e o ácido peródico de Schiff (PAS) são colorações usadas respectivamente para observar a morfologia do fígado, colágeno, fibras reticulares, gordura e glicogênio. Foram encontrados os espaços porta- hepáticos (artéria hepática, veia porta e ducto biliar e as veias centrolobulares). O glicogênio presente mostrou a média de 5,68%. O conteúdo lipídico, conferiu um aspecto goticular compatível com um quadro de esteatose hepática e a média obtida foi de 9,83%. Foi encontrado colágeno ao redor dos espaços porta-hepáticos, artérias centrolobulares e capilares sinusóides e a média foi de 14,71%. Encontrou-se também fibras reticulares ao redor dos capilares sinusóides e a média foi de 5,96%. Quanto à MET notou-se que no citoplasma dos hepatócitos desses animais existem numerosas mitocôndrias, glicogênio, muitas gotas de gordura, alguns lisossomos, retículo endoplasmático granular ao redor das mitocôndrias, algumas células estreladas, eritrócitos, núcleo, célula em degeneração e o canalículo biliar ao centro. Provavelmente o quadro sugestivo de esteatose é resultante do estado nutricional dos animais, já que nenhum outro aspecto relevante foi encontrado. Como estas aves são destinadas ao abate, sua alimentação é formulada para que os animais apresentem rápido desenvolvimento e alto ganho de peso, provavelmente com níveis de lipídios acima do necessário, causando uma deposição de micelas de gordura nos hepatócitos. Estes resultados demontraram que os hepatócitos dos avestruzes são muito simulares as outras aves apesar de também serem muito similares na estrutura das células do fígado de mamíferos. / At present the ostrich (Struthio camelus) breeding has been showing a great economical potential, although yet there are not distinct patterns about the histology of its liver, which is an organ of key importance in terms of metabolism. The knowledge of its histology and anatomy can help the detection of diseases and nutritional deficiencies that affect the growth and development of this bird. The aims of this work are to study the liver anatomical and histological structure and the branching of the hepatic artery, hepatic portal vein and bile duct. In the macroscopic study 15 ostriches with an average age of 12-18 months and average weight of 80-100 Kg, proceeding from Don Pig Abatteur, located next to Botucatu, São Paulo, were used. The birds were slaughtered with air gun and subsequently submitted to bleeding. The hepatic artery, the bile duct and the hepatic portal vein were injected with latex. The ostrich liver presents two lobules (right and left), being the right one larger than the left and both are subdivided into dorsal, intermediate and ventral. Liver samples were processed for light and electron transmission microscopic studies. Hematoxilin and eosin (HE), picrosirius, Gordon and Sweets, Sudan black and Schiff periodic acid (PAS) were respectively used to observe the liver morphology, collagen, reticular fibers, lipids and glycogen. The liver portal spaces were determined (hepatic artery, portal vein, bile duct and centrolobular veins). An average of 5.68% of glycogen was observed. The lipidic content provided a droplet aspect compatible to hepatic esteatosis, with an average of 9.83%. Collagen fibers around the liver portal spaces, centrolobular arteries and sinusoidal cappilaries were detected, at an average of 14.71%, as well as reticular fibers located in the vicinity of sinusoidal capillaries with an average of 5.96%. Through transmission electron microscopy we noticed in the hepaticyte cytoplasm the presence of numerous mithocondria, glycogen, several lipidic droplets, some lysosomes, granular endoplasmatic reticulum around the mithocondria, some stellate cells, erythrocytes, nucleus and degenerating cells, besides the central biliary canaliculus. The suggestive steatotic results might result from the animals nutritional status, once no other relevant aspect was detected. The birds are slaughtered for food comsumption and their ration is balanced striving for faster growth and higher weight gains, which are achieved through extra lipidic levels, motivating deposition of lipidic micelles in the hepatocytes. Our results demonstrated that ostrich and other birds hepatocytes are very similar.

Orientador: Márcia Rita Fernandes Machado / Banca: Tiago Luís Eilers Treichel / Banca: José Ribamar Privado Filho / Banca: Paola Castro Moraes / Banca: Fabrício Singaretti de Oliveira / Resumo: Estudou-se a anatomia do timo em 30 filhotes de avestruzes, 13 machos e 17 fêmeas com idade variando entre 10 a 20 dias, mediante fixação de cada ave em solução aquosa de formol a 10%. As cadeias de lobos tímicos localizaram-se no terço distal da região ventral do pescoço, dispondo-se de forma linear e paralela, desde a décima primeira até a décima quinta vértebra cervical, relacionando-se com o nervo vago e veia jugular. O número de lobos variou de 1 a 3 e 1 ou 2, para os antímero direito e esquerdo respectivamente. A média das dimensões dos lobos foram 2,09 x 0,71 x 0,41 cm para o lobo cranial, 0,69 x 0,38 x 0,27 cm para o lobo médio e 0,55 x 0,33 x 0,29 cm para o lobo caudal do antímero direito. No antímero esquerdo foram registrados de 2,14 x 0,71 x 0,37 cm para o lobo cranial e 0,60 x 0,31 x 0,22 cm para o lobo caudal. Os lobos tímicos direitos e esquerdos mostraram-se nutridos, em todos os casos, por vasos oriundos, direta e exclusivamente, das artérias comuns dos nervos vagos, esofágicas ascendentes e cervicais cutâneas ascendentes direitas e esquerdas. Os lobos tímicos do antímero direito receberam, independentemente de sua origem, de um a cinco ramos e do antímero esquerdo, houve uma variação de um a quatro ramos. A morfologia e vascularização arterial do timo apresentaram comportamentos bem definidos, possuindo características comuns que identificam a espécie que foi objeto de nosso estudo / Abstract: It was studied the morphology and the biometry of the thymus in 30 young ostriches, 13 males and 17 females aged from 10 to 20 days, by setting each bird in aqueous solution of 10% formaldehyde. Chains of thymic lobes were located in the third distal part of the ventral neck, offering up linear and parallel, from the eleventh to the fifteenth cervical vertebra, relating to the vagus nerve and jugular vein. The number of lobes varied from 1 to 3 and 1 or 2, to right and left antimeres respectively. The average dimensions of the lobes were 2.09 x 0.71 x 0.41 cm to the cranial lobe, 0.69 x 0.38 x 0.27 cm to the middle lobe and the 55 x 0.33 x 0.29 cm to the caudal lobe of the right side. On left antimere were found the average dimensions of the lobes of 2.14 x 0.71 x 0.37 cm for cranial lobes and 0.60 x 0.31 x 0.22 cm to the caudal lobes. Thymic right and left lobes shown to be nourished, in all cases, for vessels coming directly and exclusively, common arteries of the vagus nerves, skin and cervical esophageal ascending upward right and left . Thymic lobes of the right side received, regardless of its source, from one to five branches and left antimere, there was a variation of one to four branches. The morphology and thymus biometrics showed well-defined behaviors, possessing common characteristic who characterized the specie that was the subject of our study / Doutor

Hoje a criação de avestruz ((Struthio camelus, Linnaeus 1758)) é uma atividade de grande potencial, porém não existem padrões definidos sobre a histologia do seu fígado, que é um órgão de grande importância no metabolismo, o conhecimento de sua histologia e anatomia pode contribuir para a detecção de doenças e deficiências nutricionais que influenciam no crescimento e desenvolvimento do animal. Os objetivos desta pesquisa são: Estudar a anatomia e histologia do fígado e a ramificação de sua artéria hepática, veia porta hepática e ducto biliar. Para a realização da parte macroscópica foram utilizados quinze avestruzes com idades entre 12 e 18 meses (com peso médio em torno de 80 a 100 kg), provenientes do abatedouro Don Pig, situado próximo à cidade de Botucatu no estado de São Paulo. Os animais foram abatidos com pistola pneumática e posteriormente submetidos a sangria. Foram injetadas com látex a artéria hepática, o ducto biliar e a veia porta. O fígado dos avestruzes apresentam dois lobos (direito e esquerdo). No caso o direito é maior que o esquerdo e ambos são subdivididos em dorsal, intermédio e ventral. Além disso, amostras do fígado foram processadas para a observação em microscopia de luz e microscopia eletrônica de transmissão (MET). A hematoxilina e eosina (H.E), picrossírius, Gordon e Sweets, Sudan black e o ácido peródico de Schiff (PAS) são colorações usadas respectivamente para observar a morfologia do fígado, colágeno, fibras reticulares, gordura e glicogênio. Foram encontrados os espaços porta- hepáticos (artéria hepática, veia porta e ducto biliar e as veias centrolobulares). O glicogênio presente mostrou a média de 5,68%. O conteúdo lipídico, conferiu um aspecto goticular compatível com um quadro de esteatose hepática e a média obtida foi de 9,83%. Foi encontrado colágeno ao redor dos espaços porta-hepáticos, artérias centrolobulares e capilares sinusóides e a média foi de 14,71%. Encontrou-se também fibras reticulares ao redor dos capilares sinusóides e a média foi de 5,96%. Quanto à MET notou-se que no citoplasma dos hepatócitos desses animais existem numerosas mitocôndrias, glicogênio, muitas gotas de gordura, alguns lisossomos, retículo endoplasmático granular ao redor das mitocôndrias, algumas células estreladas, eritrócitos, núcleo, célula em degeneração e o canalículo biliar ao centro. Provavelmente o quadro sugestivo de esteatose é resultante do estado nutricional dos animais, já que nenhum outro aspecto relevante foi encontrado. Como estas aves são destinadas ao abate, sua alimentação é formulada para que os animais apresentem rápido desenvolvimento e alto ganho de peso, provavelmente com níveis de lipídios acima do necessário, causando uma deposição de micelas de gordura nos hepatócitos. Estes resultados demontraram que os hepatócitos dos avestruzes são muito simulares as outras aves apesar de também serem muito similares na estrutura das células do fígado de mamíferos. / At present the ostrich (Struthio camelus) breeding has been showing a great economical potential, although yet there are not distinct patterns about the histology of its liver, which is an organ of key importance in terms of metabolism. The knowledge of its histology and anatomy can help the detection of diseases and nutritional deficiencies that affect the growth and development of this bird. The aims of this work are to study the liver anatomical and histological structure and the branching of the hepatic artery, hepatic portal vein and bile duct. In the macroscopic study 15 ostriches with an average age of 12-18 months and average weight of 80-100 Kg, proceeding from Don Pig Abatteur, located next to Botucatu, São Paulo, were used. The birds were slaughtered with air gun and subsequently submitted to bleeding. The hepatic artery, the bile duct and the hepatic portal vein were injected with latex. The ostrich liver presents two lobules (right and left), being the right one larger than the left and both are subdivided into dorsal, intermediate and ventral. Liver samples were processed for light and electron transmission microscopic studies. Hematoxilin and eosin (HE), picrosirius, Gordon and Sweets, Sudan black and Schiff periodic acid (PAS) were respectively used to observe the liver morphology, collagen, reticular fibers, lipids and glycogen. The liver portal spaces were determined (hepatic artery, portal vein, bile duct and centrolobular veins). An average of 5.68% of glycogen was observed. The lipidic content provided a droplet aspect compatible to hepatic esteatosis, with an average of 9.83%. Collagen fibers around the liver portal spaces, centrolobular arteries and sinusoidal cappilaries were detected, at an average of 14.71%, as well as reticular fibers located in the vicinity of sinusoidal capillaries with an average of 5.96%. Through transmission electron microscopy we noticed in the hepaticyte cytoplasm the presence of numerous mithocondria, glycogen, several lipidic droplets, some lysosomes, granular endoplasmatic reticulum around the mithocondria, some stellate cells, erythrocytes, nucleus and degenerating cells, besides the central biliary canaliculus. The suggestive steatotic results might result from the animals nutritional status, once no other relevant aspect was detected. The birds are slaughtered for food comsumption and their ration is balanced striving for faster growth and higher weight gains, which are achieved through extra lipidic levels, motivating deposition of lipidic micelles in the hepatocytes. Our results demonstrated that ostrich and other birds hepatocytes are very similar.

Hierdie verhandeling is ‘n samestelling van verskeie studies, elk opgeskryf in die formaat en styl soos vereis deur die Suid-Afrikaanse Vereeniging vir Diereproduksie (South African Society of Animal Science). Hoofstuk 1 is ‘n literatuurstudie bestaande uit ‘n algemene inleiding wat agtergrond oor die studie verskaf en sluit die probleemstelling in, naamlik die swak oorlewing van volstruiskuikens wat onder kunsmatige toestande grootgemaak word. Hoofstukke 2, 3, 4 en 5 bestaan uit die eerste vier studies, waartydens sekere bestuurs-aspekte, wat moontlik oorlewing van volstruiskuikens kan verbeter (nl: populasiedigtheid, tipe vloer-oppervlak, die aanwending van broeipare en die aanwending van peetouers), ondersoek is. Hoofstukke 6, 7, 8 & 9 handel oor studies wat gerig is om voeding meer aanloklik te maak vir volstruiskuikens teneinde voedselinname te stimuleer en sodoende produksie en oorlewing te verbeter (nl: verskaffing van groenvoer, geur van voer, kleur van voer en beligting van voer). Die laaste hoofstuk bestaan uit ‘n algemene gevolgtrekking en aanbevelings. Alhoewel daar gepoog is om herhaling in hoofstukke te beperk, was dit onmoontlik om herhaling van sekere gedeeltes van hoofstukke te voorkom.

The study includes separate papers, which are all linked by their emphasis on the effect of different diets on the impact of the development of the small intestine of the young ostrich. This abstract is intended to provide readers with a broad overview of the outcomes of the study. Part 1: Yolk utilisation and the development of the small intestines Chapter 2 deals with the composition of egg yolk as it is absorbed by starved ostrich Struthio camelus L. ) chicks from one to seven days post-hatching and for ostrich (Struthio camelus L.) chicks from one to sixteen days post-hatching on a pre-starter broiler diet. Chapter 3 provides information on the intestinal enzymes of ostrich (Struthio camelus L. ) chicks from one to sixteen days post-hatching on a pre-starter broiler diet. The effect of growth on enzyme activity immediately post-hatching up to sixteen days of age was studied. Chapter 4 details a histological and morphological study of the gastrointestinal tract of ostrich (Struthio camelus L.) chicks from two to sixteen days post-hatching on a pre-starter broiler diet. The effect of growth on histological and morphological changes in the various parts of the small intestines was examined. Part 2: Influence on various pre-starter diets on growth and the development of the intestinal tract Chapter 5 discussed a growth and digestibility study of ostrich (Struthio camelus L.) chicks on eight different pre-starter diets. The performance, growth and physiological development of the chicks on high and low variations of different nutritional components were examined. Chapter 6 provides information on certain intestinal enzymes of ostrich (Struthio camelus L.) chicks on the eight different pre-starter diets. Chapter 7 dealt with the histological and morphological changes of the gastrointestinal tract of ostrich (Struthio camelus L.) chicks as a result of high and low variations of different nutritional components in prestarter diets. Implications The first part of this study revealed that although it appeared that yolk content was absorbed faster in starved ostrich chicks, intake of external feed had a positive influence on the absorption of fat from the yolk. The changes in the fatty acid composition of the yolk fat content that were observed may indicate that ostrich chicks have the ability to withdraw certain fatty acid components, especially unsaturated fatty acids. The digestive tract of ostrich chicks was compared with that of broiler chicks and differences observed in amylase and lipase activity between the birds in this trial and in poultry could be due to genetic differences between species. It could also be due to the genetic homogeneity of the poultry chicks that were used in the trials, whereas ostrich chicks have not been subjected to the same extent of advanced breeding improvement to date. Results on histological and morphological growth of the small intestines of the ostrich chick indicate increased absorption with an increase in age. The second part of this study revealed that ostrich chicks performed differently on diets formulated with different nutritional components. It appears that a high fat and low sugar content in the diet is advantageous in ostrich pre-starter diets. Although there were a few mortalities during the trials, there is no clear indication as to why the chicks died, as the causes of mortalities were not investigated. Differences in composition of the eight pre-starter diets did not seem to have an influence on the protein content or enzyme activity of the small intestines of ostrich chicks. An anti-nutritional factor within certain feedstuffs may, however, have had an influence on enzyme activity. Similar results were obtained for the histological and morphological parameters which were measured in chicks fed the different diets. There was no single diet that promoted intestine development, although it seemed as if the small intestines of the chicks fed the high protein diet were under higher digestive stress than those of the chicks fed the other diets. / Thesis (PhD)--University of Pretoria, 2011. / Anatomy and Physiology / Unrestricted

This set of experiments evaluated the physiological responses of intensively managed ostriches to L-carnitine. In experiment 1, 32 female and 16 male Zimbabwean Blue Neck and South African Black Neck breeders (n=48 of each sub-species; eight years old), were investigated in 16 breeder units of two females and one male (Trio), in a completely randomised design within four treatments and four replicates over an 8-month period during the breeding season. The same basal diet was fed supplemented with 0 (T0, control), 125 (T125), 250 (T250) or 600 (T600) mg/kg L-carnitine. T600 improved the egg production percentage, egg fertility percentage and the hatchability of set eggs for Black-Necks and Blue-Necks, respectively, and the hatchability of fertile eggs in Black Necks. L-carnitine did not affect egg shape index, defective eggs, egg weight, embryonic and post-hatch mortality. In experiment 2, 12 Black Neck males (5.5 years old) were allocated to three treatments (T0, T250 and T500) and four replicates. Semen samples were collected once a month over three months. L-carnitine had a significant effect on semen volume, sperm motility, live sperm percentage and sperm count, but had no significant effect on abnormal sperm percentage. In experiment 3, 32 day-old Black Neck ostrich chicks were allocated to treatments T0, T125, T250 and T600 with four replicates of two chicks. Chicks were vaccinated against inactive Newcastle Disease (ND) vaccine at day 30 as primary, and at day 51 as booster immunisation. ND antibody responses in the sera were monitored over three phases at 51, 70 and 80 days. Anti-NDV antibodies were detected using a modified chicken anti-NDV enzyme-like immunosorbent assay (ELISA). The treatments and the time periods and their interactions influenced ND antibody responses. T125 and T250 had the highest level of ND antibody response compare to the other groups. There were no differences in ND antibody response between T0 and T600 as well as T125 and T250. The highest ND antibody responses were recorded at day 70. Experiment 4 was designed the same as 3, to determine chicks’ growth responses over the 60-day period. Live weight and live weight gain values of T125 and T250 did not differ from those of T0. T600 had the lowest feed conversion ratio (FCR) during the total period. Feed intake (FI) was reduced in the T125 and T600 treatments compared to T0 and T250 over the total period. T125 gave the lowest FI and FCR responses over the total period, whereas there was no difference between T0 and T250. These results suggest that dietary T600 can have a beneficial effect on egg production, fertility and hatchability in the Black and Blue Neck breeders and T250 might improve sperm quality in males. In ostrich chicks T125 and T250 had positive effects on immune responses and T125 can improve the performance by decreasing the FCR. In contrast, the suppressive effect of a high inclusion level (T600) might indicate that ostrich chicks are sensitive to high inclusion levels that could cause adverse effects. / gm2013 / Animal and Wildlife Sciences / Thesis (PhD)--University of Pretoria, 2013. / Unrestricted